O.l.d'ST‘lGI/(Nil
`
`PTO/S8105 (07-O6)
`Approved for use through 01/31/2007. OMB 0651-0032
`US. Patent and Trademark Office. U.S. DEPARTMENT OF COMMEICE
`uiredto -,-i d be collection ofintormalion unless it dis avalid OMB control number.
`
`UTILITY
`
`Attomey Docket No.
`
`3‘”5 °‘”2
`Jin Po Lee
`
`PATENT APPLICATION
`TRANSMIWAL
`ronimrmwmmwsmamwmsunwrsvcsn
`
`APPLICATION ELEMENTS
`See MPEP chapter 600 wncerning Utility Patent applivafiw ¢0n1vn!S-
`
`ADDRE3s To;
`
`Commissioner for Patents
`p_o_ 3., 1450
`Alexandria VA 22313-1450
`
`1.
`
`i.
`
`Fee Transmittal Form (e.g., PTOISB/17)
`- (Submit an original and a duplicate forfee proce.w'rrg)
`2‘ I ASpe:l'§.7'(':tF"'.Rla1'Tr2':_s"'a" em"! status‘
`s
`ifi
`ti
`T tal Pa
`23
`3.
`' a'?;i.°.i;‘?i..‘:.'; and abdrad ...‘..§’.....i...?§?..T....—
`{Forinforrnation on the preferred arrangement, see MP? 608.01(a))
`Drawingls) (35 U.S.C. 113)
`[Total Sheets _6_j]
`4.
`5. Oath or Declaration
`[Total Sheets _29
`a.
`Newly exeurted (original or copy)
`b. - A copy from a prior application (37 CFR 1.63(d))
`for continuation/divisional with Box 18 completed)
`DELETION OF INVENTOR]S)
`name in the prior applimtion. see 37 CFR
`5390* Siaiemfim Efiached ‘M91309 WVBNDTKS)
`1.63(d)(2) and 1.33(b).
`
`9. El Assignment Papers (cover sheet & document(s))
`.
`Nameomss-anee
`
`10. [:| 31 CFR 3.73(b) Statement
`(when there is an assignee)
`
`Power or
`Attorney
`
`11. D English Translatlon Document (if appliwble)
`
`12.
`
`Infofltion Disclosure Statement (PTOISB/08 or PTO-1449)
`Copies of citations attached
`
`I
`
`11/650280
`
`‘UllllflllllllllllllllWHill010507
`
`6. |:| Application Data Sheet. See 37 CFR 1.76
`7.|:| CD-ROM orcD-R in duplicate, large table or
`Eputer Program (Appendix)
`Landscape Table on CD
`8. Nucleotide andlor Amlno Acld Sequence Submission
`(if ap Iicable, items a. - c. are required)
`a.
`Computer Readable Form (CRF)
`_
`S edfi
`ti
`l_- 3-
`b
`p
`ca on Sequence '5 "9 on
`i. D CD-ROM orCD-R (2 copies); or
`ii_ E]
`paper
`
`Z
`
`13_ [:1 Preliminary Mmdmm
`14
`Ram R
`. P tam MPEP5o3
`)
`' (shr¢;‘um°b°:'s':ec$iscaIryire(mizea)
`15- E]
`Copy of Priori_ty Documentlsl
`(if lbrergn pnonty rs claimed)
`,
`_
`_
`Nonpublrcation Request under 35 U.S.C. 122(b)(2)(B)(r).
`E] Applicant must attach fonn PTOISBI35 or equivalent.
`E]
`
`16.
`
`17.
`
`Other.
`
`c. D Statements verifying identity of above copies
`18. If a CONTINUING APPLICATION, check appropriate box, and supply the requisite infomiation below and in the first sentence of the
`specification following the title, or in an Application Data Sheet under 37 CFR 1. 76:
`
`El Continuation
`Prior application information.
`
`D Continuation—in-pan (CIP)
`Divisional
`Examiner LA Ajgggflgq
`
`ofprior application No.21131:1951‘)..................
`Art Unit: jlfi
`
`19. CORRESPONDENCE ADDRESS
`
`Theaddress associatedwith Customer Number:
`
`OR B Correspondenceaddress below
`
`y?-'7
`4
`/
`iI'WW/r"/////7'
`
`Email
`
`This collection at infopnation is required by 37 CFR 1.530)). The information is required to obtain or retain a benefit by the public which is to file (and by the
`USPTO to process) an application. confidentiality is governed by 35 U.S.C. 122 and 37 CFR 1.11 and 1.14. This collection is estimated to take 12 minutes to
`complete, inctuding gathering, preparing, and submitting the completed application form to the USPTO. ‘fime will vary depending upon the individual case. Any
`comments on the amount of time you require to complete this fon11 and/or suggestions for reducing this burden, should be sent to the Chief Information Officer,
`U.S. Patent and Trademark Office. U.S. Department of Commerce. P.O. Box 1450. Alexandria. VA 22313-1450. DO NOT SEND FEES OR COMPLETED
`FORMS TO THIS ADDRESS. SEND TO: Commissioner for Patents. P.O. Box 1450. Alexandria. VA 22313-1450.
`r
`Ifyou needassistance in completing the form, call 1-800-PTO43199 and ajglectoption 2.
`
`\A
`
`ALERE EX. 1002
`
`1 of 201
`
`

`
`PTOISBI17 (o7-as)
`Appxmedfor usethmugh o1I31I2n07. OMB 0851-0032
`US. Patent and Trademark Olfit»; U.S. DEPARTMBJT OF COMMERCE
`-v-tnaeol1ediaI1o1irIfctnImotILuI!emlt-v-;;.~aVafid0WlBO0fltI0l number
`
`Appncam daims small entity status. see 37 CFR 127
`
`TOTAL Amoum or Pmmazm ($)
`
`500.00
`
`Check ij Credit Card EMoney Order aNone DOther (please identify):
`Deposit Account Deposn Amoum Number:_1_9n_1_3_5________ Depodl Account Name: Bemd W. Sandt
`ForflIeabove4denfifieddepositaooomn.ttIeDiredorisherebyauflIorized to:(ched:ai1flIalapp!y)
`
`ljcIIar9efee(s)ImfIcaIe¢oeIow,exeeptforuIefiIingtoe
`I:lCharIIefee(s)indicatedbe!0w
`any
`ymen _
`‘
`under37CFR1.16and1.17
`‘/ own
`mama
`ts
`Charge any additional fee(s) or underpayments of fee(s)
`WARNING: InfommiononthisfwmnaybaemmpuNk.OednmmhfimraflonshmMmnIn|mhniodmmsfixm.Hufldeemditwm
`lnfonnatlon and aImIor|za1ion on PTO-2038»
`FEE CALCULATION
`
`EXAMINATION FEES
`smau Em
`ram &.(§)
`200
`100
`130
`65
`160
`80
`
`1. BASIC HUNG, SEARCH, AND EXAMINATION FEES
`FILING FEES
`SEARCH FEES
`Small ELI1i_ty
`Small E_nm
`M ME Fee (§)
`mm Fee (31
`Utility
`300
`150
`500
`250
`Design
`200
`100
`100
`50
`Plant
`200
`100
`300
`150
`Reissue
`300
`150
`500
`Provisional
`200
`100
`0
`2. EXCESS CLAIM FEES
`Fee Dascriflgn
`Each claim over 20 (including Reissues)
`Each independent claim ova’ 3 (including Reissues)
`Multiple dependent claims
`Fee [§)
`Toial Claims
`Extra claims
`Q
`)1
`§
`- 20 or HP =
`Q
`HP=mgImstnmmudmudaimspa5dfu.i1gra1aInan2o.
`lndeg. Clglms
`Extra Claims
`Fee (5)
`=
`it
`],
`- 3 «HP =
`1]
`I]
`HP = highest numheronnaependens dains paidfor. lfgreaterthan 3.
`3. APPLICATION SIZE FEE
`
`Fee Paid [§[
`Q
`
`=
`
`Fee Paid |§)
`0
`
`If the specification and drawings exceed 100 sheets of paper (excluding electronically filed sequence or computer
`listings under 37 CFR l.52(e)), the application size fee due is $250 ($125 for small entity) for each additional 50
`sheets or fraction thereof. See 35 U.S.C. 41 a)(1)(G) and 37 CFR 1.16 s).
`thereof
`Total Sheets
`Extra sheets
`Num
`of each additional 50 or
`23
`Q
`I50= ___0j(munduptoawhoIemnnher) x
`
`-100=
`
`Fee |§|
`0
`
`=
`
`Fg Paid ($1
`0
`
`4. OTHER FEE(S)
`Non-English Specification,
`
`$130 fee (no small entity discount)
`
`0ther_(e.g., late filinurcharge):
`
`pa“
`
`iduniafimistemfiredtoobtainorrelainabenefilbythepublicwhiehislofila(andbythe
`Thisco!!ectionofiIIformationisrequfredby37CFR1.136.'
`~-—- by35U.S.C.122and37CFR1.14. ThiscofledionisestimatedIotake30nIinutatacornplete_
`USPTOtopmcess)anappfrI:ItiurL g-- ‘—
`induding gathering. pIepa1ing,andsIIt2n1itfing1hea:Inpletedam'.i¢:afionfoImtotha USPTO. Tm1ewfllvarydepem§1IgupoIItheintfI\Iidual $9. Anycomrnents
`ontheamountoffimeyoureqIiIatnuxIIplaemisfmmandIwsuggeaionsfwIedudrIguisb:IrdeII,shoI.IIdbeseIntott:eC7iefIIIfomIa1ioII0ffieeI’.U.S.Patent
`andTrademanc0ffioe.U.S.Depa1tInemofConmIeI'ee.P.O.Bax145o,A!mIamiria,VA22313-1450. DONOTSBDFEESORCOMPLETEDFORMSTOTHIS
`ADDRESS. SEND TO: r for Patents. P.0. Box 1450. Alexandria, VA 22313-1450.
`If you need assistance in completing the form, call 1-800—PTO-9199 and seiect option 2.
`
`2 of 201
`
`

`
`11/650280
`
`‘UllllflllllllllllllllWHill010507
`
`
`
`
`
`O.l.d'ST‘lGI/(Nil
`
`PTO/S8105 (07-O6)
`Approved for use through 01/31/2007. OMB 0651-0032
`US. Patent and Trademark Office. U.S. DEPARTMENT OF COMMEICE
`uiredto -,-i d be collection ofintormalion unless it dis avalid OMB control number.
`
`UTILITY
`
`Attomey Docket No.
`
`3‘”5 °‘”2
`Jin Po Lee
`
`PATENT APPLICATION
`TRANSMIWAL
`ronimrmwmmwsmamwmsunwrsvcsn
`
`APPLICATION ELEMENTS
`See MPEP chapter 600 wncerning Utility Patent applivafiw ¢0n1vn!S-
`
`ADDRE3s To;
`
`Commissioner for Patents
`p_o_ 3., 1450
`Alexandria VA 22313-1450
`
`1.
`
`i.
`
`Fee Transmittal Form (e.g., PTOISB/17)
`- (Submit an original and a duplicate forfee proce.w'rrg)
`2‘ I ASpe:l'§.7'(':tF"'.Rla1'Tr2':_s"'a" em"! status‘
`s
`ifi
`ti
`T tal Pa
`23
`3.
`' a'?;i.°.i;‘?i..‘:.'; and abdrad ...‘..§’.....i...?§?..T....—
`{Forinforrnation on the preferred arrangement, see MP? 608.01(a))
`Drawingls) (35 U.S.C. 113)
`[Total Sheets _6_j]
`4.
`5. Oath or Declaration
`[Total Sheets _29
`a.
`Newly exeurted (original or copy)
`b. - A copy from a prior application (37 CFR 1.63(d))
`for continuation/divisional with Box 18 completed)
`DELETION OF INVENTOR]S)
`name in the prior applimtion. see 37 CFR
`5390* Siaiemfim Efiached ‘M91309 WVBNDTKS)
`1.63(d)(2) and 1.33(b).
`
`9. El Assignment Papers (cover sheet & document(s))
`.
`Nameomss-anee
`
`10. [:| 31 CFR 3.73(b) Statement
`(when there is an assignee)
`
`Power or
`Attorney
`
`11. D English Translatlon Document (if appliwble)
`
`12.
`
`Infofltion Disclosure Statement (PTOISB/08 or PTO-1449)
`Copies of citations attached
`
`I
`
`6. |:| Application Data Sheet. See 37 CFR 1.76
`7.|:| CD-ROM orcD-R in duplicate, large table or
`Eputer Program (Appendix)
`Landscape Table on CD
`8. Nucleotide andlor Amlno Acld Sequence Submission
`(if ap Iicable, items a. - c. are required)
`a.
`Computer Readable Form (CRF)
`_
`S edfi
`ti
`l_- 3-
`b
`p
`ca on Sequence '5 "9 on
`i. D CD-ROM orCD-R (2 copies); or
`ii_ E]
`paper
`
`Z
`
`13_ [:1 Preliminary Mmdmm
`14
`Ram R
`. P tam MPEP5o3
`)
`' (shr¢;‘um°b°:'s':ec$iscaIryire(mizea)
`15- E]
`Copy of Priori_ty Documentlsl
`(if lbrergn pnonty rs claimed)
`,
`_
`_
`Nonpublrcation Request under 35 U.S.C. 122(b)(2)(B)(r).
`E] Applicant must attach fonn PTOISBI35 or equivalent.
`E]
`
`16.
`
`17.
`
`Other.
`
`c. D Statements verifying identity of above copies
`18. If a CONTINUING APPLICATION, check appropriate box, and supply the requisite infomiation below and in the first sentence of the
`specification following the title, or in an Application Data Sheet under 37 CFR 1. 76:
`
`El Continuation
`Prior application information.
`
`D Continuation—in-pan (CIP)
`Divisional
`Examiner LA Ajgggflgq
`
`ofprior application No.21131:1951‘)..................
`Art Unit: jlfi
`
`19. CORRESPONDENCE ADDRESS
`
`Theaddress associatedwith Customer Number:
`
`OR B Correspondenceaddress below
`
`y?-'7
`4
`/
`iI'WW/r"/////7'
`
`Email
`
`This collection at infopnation is required by 37 CFR 1.530)). The information is required to obtain or retain a benefit by the public which is to file (and by the
`USPTO to process) an application. confidentiality is governed by 35 U.S.C. 122 and 37 CFR 1.11 and 1.14. This collection is estimated to take 12 minutes to
`complete, inctuding gathering, preparing, and submitting the completed application form to the USPTO. ‘fime will vary depending upon the individual case. Any
`comments on the amount of time you require to complete this fon11 and/or suggestions for reducing this burden, should be sent to the Chief Information Officer,
`U.S. Patent and Trademark Office. U.S. Department of Commerce. P.O. Box 1450. Alexandria. VA 22313-1450. DO NOT SEND FEES OR COMPLETED
`FORMS TO THIS ADDRESS. SEND TO: Commissioner for Patents. P.O. Box 1450. Alexandria. VA 22313-1450.
`r
`Ifyou needassistance in completing the form, call 1-800-PTO43199 and ajglectoption 2.
`
`3 of 201
`
`

`
`PTOISBI17 (o7-as)
`Appxmedfor usethmugh o1I31I2n07. OMB 0851-0032
`US. Patent and Trademark Olfit»; U.S. DEPARTMBJT OF COMMERCE
`-v-tnaeol1ediaI1o1irIfctnImotILuI!emlt-v-;;.~aVafid0WlBO0fltI0l number
`
`Appncam daims small entity status. see 37 CFR 127
`
`TOTAL Amoum or Pmmazm ($)
`
`500.00
`
`Check ij Credit Card EMoney Order aNone DOther (please identify):
`Deposit Account Deposn Amoum Number:_1_9n_1_3_5________ Depodl Account Name: Bemd W. Sandt
`ForflIeabove4denfifieddepositaooomn.ttIeDiredorisherebyauflIorized to:(ched:ai1flIalapp!y)
`
`ljcIIar9efee(s)ImfIcaIe¢oeIow,exeeptforuIefiIingtoe
`I:lCharIIefee(s)indicatedbe!0w
`any
`ymen _
`‘
`under37CFR1.16and1.17
`‘/ own
`mama
`ts
`Charge any additional fee(s) or underpayments of fee(s)
`WARNING: InfommiononthisfwmnaybaemmpuNk.OednmmhfimraflonshmMmnIn|mhniodmmsfixm.Hufldeemditwm
`lnfonnatlon and aImIor|za1ion on PTO-2038»
`FEE CALCULATION
`
`EXAMINATION FEES
`smau Em
`ram &.(§)
`200
`100
`130
`65
`160
`80
`
`1. BASIC HUNG, SEARCH, AND EXAMINATION FEES
`FILING FEES
`SEARCH FEES
`Small ELI1i_ty
`Small E_nm
`M ME Fee (§)
`mm Fee (31
`Utility
`300
`150
`500
`250
`Design
`200
`100
`100
`50
`Plant
`200
`100
`300
`150
`Reissue
`300
`150
`500
`Provisional
`200
`100
`0
`2. EXCESS CLAIM FEES
`Fee Dascriflgn
`Each claim over 20 (including Reissues)
`Each independent claim ova’ 3 (including Reissues)
`Multiple dependent claims
`Fee [§)
`Toial Claims
`Extra claims
`Q
`)1
`§
`- 20 or HP =
`Q
`HP=mgImstnmmudmudaimspa5dfu.i1gra1aInan2o.
`lndeg. Clglms
`Extra Claims
`Fee (5)
`=
`it
`],
`- 3 «HP =
`1]
`I]
`HP = highest numheronnaependens dains paidfor. lfgreaterthan 3.
`3. APPLICATION SIZE FEE
`
`Fee Paid [§[
`Q
`
`=
`
`Fee Paid |§)
`0
`
`If the specification and drawings exceed 100 sheets of paper (excluding electronically filed sequence or computer
`listings under 37 CFR l.52(e)), the application size fee due is $250 ($125 for small entity) for each additional 50
`sheets or fraction thereof. See 35 U.S.C. 41 a)(1)(G) and 37 CFR 1.16 s).
`thereof
`Total Sheets
`Extra sheets
`Num
`of each additional 50 or
`23
`Q
`I50= ___0j(munduptoawhoIemnnher) x
`
`-100=
`
`Fee |§|
`0
`
`=
`
`Fg Paid ($1
`0
`
`4. OTHER FEE(S)
`Non-English Specification,
`
`$130 fee (no small entity discount)
`
`0ther_(e.g., late filinurcharge):
`
`pa“
`
`iduniafimistemfiredtoobtainorrelainabenefilbythepublicwhiehislofila(andbythe
`Thisco!!ectionofiIIformationisrequfredby37CFR1.136.'
`~-—- by35U.S.C.122and37CFR1.14. ThiscofledionisestimatedIotake30nIinutatacornplete_
`USPTOtopmcess)anappfrI:ItiurL g-- ‘—
`induding gathering. pIepa1ing,andsIIt2n1itfing1hea:Inpletedam'.i¢:afionfoImtotha USPTO. Tm1ewfllvarydepem§1IgupoIItheintfI\Iidual $9. Anycomrnents
`ontheamountoffimeyoureqIiIatnuxIIplaemisfmmandIwsuggeaionsfwIedudrIguisb:IrdeII,shoI.IIdbeseIntott:eC7iefIIIfomIa1ioII0ffieeI’.U.S.Patent
`andTrademanc0ffioe.U.S.Depa1tInemofConmIeI'ee.P.O.Bax145o,A!mIamiria,VA22313-1450. DONOTSBDFEESORCOMPLETEDFORMSTOTHIS
`ADDRESS. SEND TO: r for Patents. P.0. Box 1450. Alexandria, VA 22313-1450.
`If you need assistance in completing the form, call 1-800—PTO-9199 and seiect option 2.
`
`4 of 201
`
`

`
`MULTIPLE ANALYTE ASSAY DEVICE
`
`FIELD OF THE INVENTION
`
`5 The present invention relates to methods and devices for assaying biological fluid
`
`samples. More particularly, the invention relates to methods and devices for detecting
`
`analytes, such as drugs, in urine. The present application is a division of copending
`
`application SN 10/ 01 90570 filed November 8, 2001.
`
`HISTORY OF THE RELATED ART
`
`In their most simple form, chromatographic analyte test strips permit an assay to be
`
`10 performed in a single step (application of an analyte sample to the device) to produce
`
`visually observable assay results (such as those indicated by colored bars on the test
`
`strip). However, a common limitation of such test strips is that they can only be used to
`
`detect a single analyze, requiring that serial assay procedures be performed to detect
`
`additional analytes (for example, to test a sample for the presence of a panel of
`
`15 narcotics). Multiple dipping steps, such as are commonly used when multiple dipstick
`
`assays are separately performed, present not only a possible loss of sensitivity of the
`
`assay (through reagent mixing or loss of reagent solutions), but also an esthetic and
`
`hygienic problem for the analyst. Repetitive performance of assay procedures is also
`
`tedious, which increases the risk that assays will be performed improperly or the results
`
`20 misinterpreted.
`
`5 of 201
`
`

`
`A‘ W0 00/05579
`
`-
`
`'
`
`I
`
`’
`
`PCT/US98/15369
`
`-2-
`
`SUMMARY OF THE lNVEl\lTIO.N
`
`The present invention provides an assay device, device for separating a fluid analyte
`
`sample for use in multiple assay procedures and methods for performing multiple analyte
`
`assays.
`In one embodiment of the assay device, the assay device is a dipstick having
`multiple analyte test strips, each of which includes atest zone and a control zone. The
`
`test strips are enclosed in a housing having an open side through which an end of each
`
`test strip protrudes to form a sample loading zone. A protective cap is provided to seal
`
`the protruding ends of the test strips from exposure while not in use. Each test strip is
`separated from the next within the housing by a raised spacer. The portion ofthe housing
`which overlies the test and control zones is transparent to permit visually observible
`results shown in each zone to be viewed.
`
`In cassette form, the assay device has the same structure described above, but the
`
`protruding test strips are inserted into a cap which has a sample port for application of
`
`sample to the test strips. The cap is retained on the assay device by a close fit over the
`
`device housing.
`
`Each test strip provides binders and assay reagents for detection of a different analyte in
`
`the sample fluid. In a particularly preferred embodiment of the assay device, the housing
`
`may be opened to permit substitution of different test strips to allow each device to be
`
`customized for detection of specific analytes of interest. Assay sample integrity
`
`determinants consisting of test strips which allow measurement of parameters such as
`
`specific gravity and pH may also be included in each device.
`
`The invention also provides a separator device for dividing a fluid assay sample into
`
`portions for use in multiple assays without need for contact between the assay operator
`
`and the fluid sample. This latter feature of the device increases operator safety and
`
`avoids inadvertent contamination of the assay sample. The separator device may be used
`
`to separate any fluid assay sample, but is especially useful in assaying samples for the
`
`6 of 201
`
`

`
`W000/05579
`
`‘
`
`.
`
`'
`
`i
`
`i
`
`‘
`
`PCT/US98/15369
`
`-3-
`
`presence of narcotics, where a "positive" result on first testing of the sample may
`
`necessarily be followed by additional testing of the sample to confirm the result and the
`
`identity of the detected narcotic. To this end, the separator device is adapted particularly
`
`well to use with the assay ‘device of this invention.
`
`The assay device of the invention makes specimen analysis easier because an analyte
`
`sample need only to be applied once to the assay device for testing. In addition, the
`
`replaceable nature of the analyte test strips allows the analyst to customize the array of
`
`assays to the testing situation. Because the customization can be performed before adding
`
`the test sample (e.g., urine), fewer manipulations with the analyte sample are needed to
`obtain the desired information.
`In addition, use of the separator device permits further
`
`testing of the sample to be performed without risk of adultering the sample in a
`
`preliminary assay performed according to the invention.
`
`The invention also provides a method for assaying one or more analytes of interest. The
`
`protruding ends of the device are dipped into a fluid analyte sample. Binding of an
`
`analyte present in the sample with one or more specific ligands causes formation of
`
`specific visual pattern in the test and control zones indicative of the test result. The assay
`
`results performed according to the invention may be read visually without use of separate
`
`measuring equipment. Thus, performance of assays according to the invention requires
`
`only that the user introduce the requisite amount of test sample into the device of the
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`invention, then observe any color changes which appear shortly thereafter in a detection
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`zone of an analyte strip. The method of the invention is especially useful for screening
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`fluid analyte samples (e.g., urine) for the presence or absence of drugs of abuse.
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`BRIEF DESCRIPTION OF‘ THE DRAWINGS
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`FIGURE 1 is an exploded view of a dipstick assay device of the invention.
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`FIGURE 2 is atop view of a dipstick assay device of the invention.
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`FIGURE 3A is a top view of the upper half sample port cap of a cassette assay device of
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`the invention, and FIGURE 3B is a top view of the lower half, base portion of the sample
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`port cap, while FIGURE 3C is a side, cut-away view of the intact cap with test strips in
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`place therein.
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`FIGURE 4 is a top view of the separator device of the invention.
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`FIGURE 5 is a cross—sectional view of the separator device taken along line A—A at cut-
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`away point B-B ofFIGURE
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`FIGURE 6 is a lateral view of the separator device within a specimen collection cup.
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`Like numerals refer to like elements in the drawings.
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`DETAILED DESCRIPTION OF INVENTION
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`A.
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`Definitions
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`For ease of understanding,
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`the following definitions will apply throughout
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`this
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`description:
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`I.
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`The term "antigen" as used herein refers to any analyte which is
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`capable of binding antibodies. Antigens may comprise, without limitation, chemical
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`compounds, polypeptides, carbohydrates, nucleic acids, lipids, and the like, including
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`viral particles, viral subunits, bacterial and parasite surface antigens, and host proteins
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`that may be diagnostic of the subject's condition.
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`2.
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`A "binder" refers to a ligand for the analyte as in the format of a
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`sandwich assay, or a ligand for both the analyte and the tracer as in the format of a
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`competitive assay. A binder can be chosen from a group of molecules or compounds
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`capable of binding the analyte, such as an antigen to the antibody analyte, or an antibody
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`to the antigen analyte.
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`3.
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`A “test zone” refers to an area in which a binder or the analyte is
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`attached, movably or immovably, to the test strip portion of an assay device.
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`4.
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`A "tracer" refers to a ligand for the analyte or the binder labeled with
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`a detectable label, preferably a visually readable particulate label, such as colloidal gold,
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`latex and liposomes including dye, carbon black, and the like.
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`5.
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`A "sample loading zone" refers to an area of a test strip on which a
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`fluid analyte sample is applied for migration to the test zone.
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`6.
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`A “test strip” of the invention consists of, collectively, all of the zone
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`supporting membranes and any filters of the assay device.
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`7.
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`A “fluid analyte sample” can be any fluid suspected of containing
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`analyte ofinterest for which a particular assay is specific. Test sample may represent any
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`body fluid, including urine, blood, sweat, lymph, intraperitoneal fluid, crude tissue
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`extract or homogenate, derived from a fetus, neonate, juvenile or adult subject; a non-
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`biological fluid such as water from some ecological niche, e.g., a river or a lake; or a
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`solution used in a laboratory.
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`8.
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`A “label” is a molecule or compound which directly or indirectly
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`mediates the formation of a signal (such as a color change) which is used in assay to
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`indicate the presence, absence or concentration range of analyte of interest in a test-
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`sample. Labels may include enzymes, fluorescers,
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`liposomes, erythrocyte ghosts,
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`polymer microcapsules, color polymer particles (latex), and preferably includes sols of
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`metal-containing compounds. A wide variety of patents and patent applications provide
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`an extensive literature of different
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`techniques for producing detectible signals in
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`immunoassays. The following list of United States patents is merely illustrative of the
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`type of label which can find application in this invention: U.S. Patent Nos. 3,646,346
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`discloses radioactive label; 3,654,090, 3,791,932, and 3,817,838 disclose enzyme labels;
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`3,996,345 discloses fluorescer—quencher labels; 4,062,733 discloses radioactive label;
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`4,067,959 discloses fluorescer or enzyme label; 4,104,099 discloses chemiluminescent
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`1abel;‘and 4,160,645 discloses non-enzymatic catalyst label. U.S. Patent No. 3,966,879
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`discloses an electrophoretic technique employing an antibody zone and U.S. Patent No.
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`4,120,945 discloses a radioimmune assay (RIA) where labeled analyte is initially bound
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`to a solid support through antibody. U.S. Patent No. 4,233,402 discloses enzyme pair
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`labels; U.S. Patent No. 4,720,450 discloses chemically induced fluorescent labels; and
`U.S. Patent No. 4,287,300 discloses enzyme anionic charge labels.
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`Labels can also be metal-containing sols; i.e., metal or metal compounds such
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`as metal oxides, metal hydroxides, metal salts, metals or metal-containing compounds
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`mixed with polymers or coated onto polymer nuclei. These metal labels may include dry
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`forms of any ofthe above-named metal or metal compound sols, and preferably includes
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`colloidal gold in dry form.
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`9.
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`A “complex” means (depending on the context) any multimolecular
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`complex formed by analyte and one or more ligands, or by labeled ligand and
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`immobilized ligand. In a sandwich-type immunoassay, e.g., the following complexes
`occur: analyte/labeled ligand duplex first produced in the assay (first complex) and
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`analyte/labeled ligand/immobilized ligand triplex formed second in the assay (second
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`complex).
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`10.
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`“Fluid communication” refers to structures which are in contact with,
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`but not necessarily affixed to, one another.
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`11.
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`“Assay” refers to severaldifferent types of assay formats in which an
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`analyte of interest can be detected using an assaytest strip. For example, in a sandwich-
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`type immunoassay, analytes of interest in the analyte sample, when present, bind a
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`labeled tracer movably incorporated in the test strip (consisting of a porous membrane)
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`at the tracer zone to form a first complex. The tracer is a molecule which binds the
`analyte of interest and is conjugated to a label, preferably a metal label, and most
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`preferably colloidal gold.
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`A second immobilized ligand corresponding to the analyte of interest is
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`coupled to the test strip at the test zone. First complex and unbound labeled ligand mix
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`with the test sample and be carried along therewith by capillary action (wicking) through
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`the test zone. Analyte sample passes through the test strip bringing the first complexes,
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`if any, into contact with the unlabeled ligand immobilized in the test zone to form a
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`second complex of labeled ligand-analyte-immobilized ligand. The first immobilized
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`ligand is immobilized in the test zone by means known in the art, including covalent
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`bonding or attachment to an insoluble protein-coated surface (see, e.g., U.S. Patents No.
`4,200,690 and 5,075,078). When the second complex is formed, a visible color pattern
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`appearspin the test zone. Labeled ligand not bound to analyte in the test sample continue
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`migration by wicking into the control zone to contact the ligand immobilized there.. The
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`labeled ligand can bind the immobilized ligand in the control zone to form a third
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`complex, and thus be captured in the control zone.
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`Within the scope ofthis invention, the labeled ligand formingithe complex in
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`the control zone may be the same as the tracer forming the first and second complexes,
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`or it may be a different labeled ligand. The ligand immobilized in the control zone should
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`have specific affinity for the labeled ligand intended to form the third complex.
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`Formation of the third complex is indicated by a visible pattern in the control zone.
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`Besides sandwich immunoassay method, other assay methods may be
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`implemented in the devices of the invention. These methods may include competition
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`and inhibition assays. In a competition assay, the analyte and tracer have similar affinity
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`properties and compete for binding with immobilized ligand. Thus, in absence of analyte,
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`the pattern (e.g., band) in the test zone is of maximum intensity. When present, the
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`analyte binds to immobilized ligand to prevent the tracer from getting captured in the test
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`zone. Thus, the intensity of the test band is reduced, depending on the concentration of
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`analyte in the test sample.
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`In an inhibition assay, the analyte and immobilized ligand in the test zone
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`each have affinity for the tracer. In the absence of analyte in the analyte sample, the
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`tracer is captured by immobilized ligand, and a visible pattern forms in the test zone.
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`When present, the analyte binds the tracer, thereby preventing it from binding to the
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`immobilized ligand in the test zone. The resulting intensity of the test band is reduced
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`depending on the concentration of analyte in the test sample.
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`Dipstick Assay Device
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`Turning to FIG. 1, a dipstick form of the assay device is shown in exploded view. The
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`device consists of a housing 100, which is defined by base 101 and cover 110. Base 101
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`can be constructed of any sterilizable material, such as a nonporous plastic (e.g., the
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`commercially available plastic “ABS” supplied by the Monsanto Company of St. Louis,
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`MO.). Base 101 having a closed end 104 and an open end 106, slots 102A, 102B, 102C,
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`102D and 102E separated by rails 103A, 103B, 103C and 103D for insertion of test strips
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`105A, 105B, 105C, 105D and 105E. A particular advantage of this embodiment of the
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`assay device is its customizability in that test strips specific for different analytes of
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`interest to the user may be inserted into base 101 and that the number of test strips
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`employed may vary (e.g., base 101 may have any number of slots from two upward to
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`accomodate as many test strips as the user may desire).
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`Referring to FIG. 2, when inserted into slots 102A, 102B, 102C, 102D and 102E, the test
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`strips extend out of base 101 beyond open end 106. The length to which the test strips
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`protrude from base 101 must be sufficient to allow the test strips to contact a fluid
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`analyte sample, preferably by immersion, and most preferably without allowing the fluid
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`to contact housing 100. The test strips are conventional in form; therefore, because those
`ofordinary skill in the art will be abundantly familiar with the design of such test strips,
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`they will not be described in detail here. However, each test strip will have a test zone
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`112 for binding of analyte (to indicate a positive test result for the presence of analyte in
`the analyte sample) and a control zone 113 for binding of tracer (to indicate correct
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`operation ofthe assay). Preferably, the test zones and control zones of each test strip lie
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`in the same location on each test strip so each can be viewed in side-by-side fashion.
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`Each test strip is typically constructed of a porous membrane which is substantially inert
`with respect to the analyte and must be porous or absorbent relative to the analyte sample
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`to be tested, e.g., urine. The substance can be either bibulous matrices or nonbibulous.
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`matrices that are insoluble in, and maintain their structural integrity when exposed to
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`aqueous solutions or'physiological.fluids. Bibulous matrices that can be useful for the
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`devices of the present invention include but are not limited to, paper, sponge materials,
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`cellulose, hydrophilic inorganic powders, wood, synthetic resin fleeces, woven and
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`nonwoven fabrics and like materials. Nonlimiting examples of nonbibulous matrices
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`include glass fiber, penneable polymer films and preformed or microporous membranes.
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`The absorbent material is preferably absorbent paper. The absorbent material can be
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`affixed by a double sided adhesive (e.g., two sided adhesive tape) to a solid moisture
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`impervious support. This support can be constructed from, for example, hydrophobic
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`plastic, cellulose acetate, polyethylene, terephthalate, polycarbonate, or polystyrene.
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`The tracer is prepared according to the means known in the art. For purposes of
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`producing a clearly visible reaction, labels of metal—containing sols are preferred, with
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`labels of colloidal gold or selenium being most preferred. An example of a suitable
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`product is colloidal gold available from Janssen Life Sciences Products. These colloidal
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`metals produce distinctive visual patterns without addition of further reagents; however,
`fluorescers (such as fluorescein) and enzymes (such asthose identified in U.S. Patent No.
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`4,275,149), may also be used.
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`Selections and choices for test binders (e.g., immobilized antigens, antibodies and other
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`test and control binders), as well as suitable means for their attachment to porous test
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`strip membranes, are well-known to those of ordinary skill in the art and will not be
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`stated in detail here. To maximize contact of test sample with the tracer and all test
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`binders, the area occupied by each reagent on the test strip preferably extends from one
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`side of the membrane to the other.
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`For further review concerning test strip construction, including selection and preparation
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`of test reagents, the following references provide a representative sample of test strip
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`designs known in the art: US Patent No. 5,3 84,264 (commonly owned); US Patent No.
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`4,491,645; US Patent No. 4,943,522; US Patent No. 5,252,496; US Patent No. 5,714,389
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`and US Patent No. 5,602,040, the disclosures of which are incorporated for purposes of
`reference.
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`Test strips 105A, 105B, 105C, 1051) and 105E may be secured within slots 102A, 102B, ‘
`102C, 102D and 102E by adhesion to the floor of each slot; however, the placement of
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`cover 110 onto base 101 is sufficient to retain the test strips within the base s