(12) United States Patent
`Cabilly et al.
`
`(10) Patent No.:
`
`(45) Date of Patent:
`
`US 6,331,415 B1
`Dec. 18, 2001
`
`US006331415B1
`
`(54) METHODS OF PRODUCING
`IMMUNOGLOBULINS, VECTORS AND
`TRANSFORMED HOST CELLS FOR USE
`THEREIN
`
`(75)
`
`Inventors: Shmuel Cabilly, Monrovia; Herbert L.
`Heyneker, Burlingame; William E.
`Holmes, Pacifica; Arthur D. Riggs, La
`Verne; Ronald B. Wetzel, San
`Francisco, all of CA (US)
`
`(73) Assignee: Genentech, Inc., South San Francisco,
`CA (US)
`
`( * ) Notice:
`
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 0 days.
`
`(21) Appl. No.: 07/205,419
`
`(22)
`
`Filed:
`
`Jun. 10, 1988
`
`Related U.S. Application Data
`
`(63) Continuation of application No. 06/483,457, filed on Apr. 8,
`1983, now Pat. No. 4,816,567.
`
`(51)
`
`Int. Cl.7 ......................... .. C12N 15/13, C12N 15/00,
`C12N 15/63
`
`(52) U.s. Cl.
`
`..................... .. 435/69.6; 435/691; 435/69.7;
`435/701, 435/70.21; 435/711, 435/712,
`435/320, 435/252.1, 435/252.3, 435/252.33,
`435/254.11, 435/254.2, 435/254.21; 435/455,
`435/471, 435/483; 435/485
`
`(58) Field of Search ................................ .. 435/69.1, 69.7,
`435/71.1, 70.1, 71.2, 320, 261, 252.1, 252.3,
`81, 55, 56, 69.6, 252.33, 254.21, 483
`
`(56)
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`.
`
`12/1979 Davis et al.
`4,179,337
`12/1980 Cohen .
`4,237,224
`7/1982 Gilbert et al..
`4,338,397
`8/1982 Goeddel et al. .
`4,342,832
`9/1983 Hartley et al.
`.
`4,403,036
`4/1984 Paulus.
`4,444,878
`4/1985 Parks et al. .
`4,510,244
`4/1985 Jones et al. .
`4,512,922
`5/1985 Mark et al. .
`4,518,584
`2/1987 Moore et al.
`4,642,334
`11/1987 Itakura et al.
`4,704,362
`3/1989 Boss et al.
`.
`4,816,397
`3/1989 Cabilly et al.
`4,816,567
`7/1993 Winter.
`5,225,539
`8/1996 Page .
`5,545,403
`8/1996 Page .
`5,545,404
`8/1996 Page.
`5,545,405
`FOREIGN PATENT DOCUMENTS
`
`.
`
`.
`
`.
`
`194982
`12417/83
`B—26429/84
`46556/85
`65981/86
`0037723
`
`2/1983 (AU) .
`9/1983 (AU) .
`10/1984 (AU).
`3/1986 (AU).
`5/1987 (AU).
`10/1981
`
`37723
`041313
`041767
`41313
`41767
`044722
`055945
`57107
`0068763
`68763
`0057107
`0073656
`075444
`73656
`75444
`A-073656
`088994
`88994
`093619
`0120694
`0125023
`194276
`196864
`234592
`255694
`324162
`550400
`08235
`62 201 581
`WO 86/01533
`
`10/ 981 (312).
`12/981 (312).
`12/981 (312).
`12/981 (312).
`12/981 (312).
`1/ 982 (312).
`7/ 982 (312).
`8/ 982 (312).
`1/ 983 (312).
`1/983 (312).
`3/983 (312).
`3/ 983 (312).
`3/ 983 (312).
`3/983 (312).
`3/983 (312).
`3/983 (312).
`9/983 (312).
`9/983 (312).
`11/ 983 (312).
`10/ 984 (312).
`11/ 984 (312).
`9/ 986 (312).
`10/986 (312).
`9/ 987 (312).
`2/988 (312) .
`7/ 989 (312).
`7/ 993 (312).
`3/ 987 (GB).
`9/ 987 (JP) .
`3/ 986 (W0).
`OTHER PUBLICATIONS
`
`Dolby et al. Prac. Natl. Acad. Sci. 77(10):6027—6031
`(1980).
`Rice et al. Prac. Natl. Acad. Sci. 77:7862-7865 (1982).
`Accolla et al. Proc. Natl. Acad. Sci. 77(1):563—566 (1980).
`Raso et al. Cancer Res. 41:2073—2078 (1981).
`Nisonoff et al. Arch. Biachem. Biaphys. 93:460-462 (1960).
`Glennie et al. Nature 295:712—714 (1982).
`Eisen Immunology Harper & Row, Publishers, pp. 415 and
`428-436 (1974).
`Ilozumi et al. Nuc. Acids. Res. 5(6):1779—1799 (1978).
`Wetzel et al. Gene 16:63-71 (1981).
`Williams et al. Science 215:687-689 (1982).
`Falkner et al. Nature 298:286—288 (1982).
`Boss et al. Gene Expressions-1-’r0c. Cetus-UCLA Sympo-
`sium pp. 513-522, Mar. 26-Apr. 1, 1983.
`
`(List continued on next page.)
`
`Primary Examiner—Phillip Gambel
`(74) Attorney, Agent, or Firm—Burns, Doane, Swecker &
`Mathis, LLP
`
`(57)
`
`ABSTRACT
`
`The invention relates to processes for producing an immu-
`noglobulin or an immunologically functional immunoglo-
`bulin fragment containing at least the variable domains of
`the immunoglobulin heavy and light chains. The processes
`can use one or more vectors which produce both the heavy
`and light chains or fragments thereof in a single cell. The
`invention also relates to the vectors used to produce the
`immunoglobulin or fragment, and to cells transformed with
`the vectors.
`
`36 Claims, 19 Drawing Sheets
`
`Genzyme Ex. 1001, pg 1
`
`Genzyme Ex. 1001, pg 1
`
`

`
`US 6,331,415 B1
`Page 2
`
`OTHER PUBLICATIONS
`
`Amster et al. NucleicAcid Research 8(9):2055-2065 (1980).
`DeBoer et al., Rodriguez et al. (Ed.) Promoters 462-481
`(1982).
`Gough Tibs 6(8)2203-205 (Aug., 1981).
`Morrison J. of Immunology 123(2)2793-800 (Aug., 1979).
`Kohler Proc. Natl. Acad. Sci. 77(4):2197-2199 (Apr, 1980).
`Roberts Promoters 452-461 (1982).
`Kemp et al. Proc. Natl. Acad. Sci. 78(7)24520-4524 (Jul.,
`1981).
`Valle et al. Nature 300271-74 (Nov. 4, 1982).
`Microbiology 3rd edition, Harper Int. Ed. 338-379 (1980).
`Hitzeman et al. Science 2192620-625 (1983).
`Mercereau-Puij alon et al. in Expression of Eukaryotic Viral
`and Cellular Genes, Pettersson et al. (ED) 295-303 (1981)
`Academic Press.
`
`295-303 (1981) Academic Pr.
`Pettersson et al.
`Keshet et al. Nucleic Acids Res. 9(1):19-30 (1981).
`Taniguchi et al. Proc. Natl. Acad. Sci. 77(9)25230-5233
`(1980).
`Ohsuye et al. Nucleic Acids Res. 11(5)21283-1295 (1983).
`Kadonaga et al. J. Biol. Chem. 259(4)22149-2154 (1984).
`Maniatis Molecular Cloning p. 433 (Sep., 1985).
`Fujisawa et al.NucleicAcids Res. 11(11):3581-3591 (1983).
`Roberts Promoters Structures and Function Rodriguez, R.
`L. (Ed.) 452-461 (1982).
`Adams et al. Biochemistry 1922711-2719 (1980).
`Haley et al. DNA 12155-162 (1982).
`Gough et al. Biochemistry 19:2702-2710 (1980).
`Iserentant et al. Gene 921-12 (1980).
`Seidman et al. Nature 271:582-585 (1978).
`Stevens et al. J. Mol. Biol. 782517-525 (1973).
`Deacon et al. Biochem. Soc. Trans. 4:818-820 (1976).
`Colman et al. Cell 172517-526 (1979).
`Valle et al. Nature 2912338-340 (1981).
`Cohnan et al. J. Mol. Biol. 1602459-474 (Sep., 1982).
`Cowan et al. J. Mol. Biol. 902691-701 (1974).
`Morrison et al. J. Immunol. 1142655-659 (1975).
`Mosmann et al. J. Immunol. 115:955-962 (1975).
`Levy et al. Proc. Nat. Acad. Sci. 7522411-2415 (1978).
`Robertson et al. Nature 2872390-392 (1980).
`Mosmann et al. Cell 202283-292 (1980).
`Wilde et al. Eur. J. Immunol. 102462-467 (1980).
`Ochi et al. Nature 3022340-342 (Mar. 24, 1983).
`Kemp et al. in Nagley et al., Ed., Manipulation and Expres-
`sion of Genes in Eukaryotes Proceedings of an International
`Conference, 12th International Congress of Biochemistry, in
`Australia Aug. 9-13, 1982 (1983).
`Picard et al. Proc. Nat. Acad. Sci. 802417-421 (Jan., 1983).
`Oi et al. Proc. Nat. Acad. Sci. 80:825-829 (Eeb., 1983).
`Ilawley et al. Proc. Nat. Acad. Sci. 7927425-7429 (Dec.,
`1982).
`Boss et al. Immunology Today 6(1)212-13 (1985).
`Lewin, Ed. Genes 3rd Edition, 359-360 (1987).
`Skerra et al. Science 24021038-1040 (1988).
`Kohler et al. Eur. J. Immunol. 62292-295 (1976).
`Adair et al., “Engineering Antibodies for Therapy” Immu-
`nological Reviews 13025-40 (1992).
`Adelman et al., “In Vitro Deletional Mutagenesis for Bac-
`terial Production of the 20,000-Dalton Form of Human
`Pituitary Growth Hormone” DNA 2(3):183-193 (1983).
`Adetugbo, K., “Spontaneous Somatic Mutations” Journal of
`Biological Chemistry 253(17)26076-6080 (1978).
`
`Alt et al., “Activity of Multiple Light Chain Genes in Murine
`Myeloma Cells Producing a Single, Functional Light Chain”
`Cell 2121-12 (Aug. 1980).
`Alt et al., “Joining of immunoglobulin heavy chain gene
`segments: Implications from a chromosome with evidence
`of
`three D-JH fusions” Proc. Natl. Acad. Sci. USA
`7924118-4122 (Jul. 1982).
`Alt et al., “Multiple Immunoglobulin Heavy-Chain Gene
`Transcripts in Abelson Murine Leukemia Virus-Trans-
`ormed Lymphoid Cell Lines” Molecular & Cellular Biol-
`ogy 2(4)2386-400 (Apr. 1982).
`Alt et al., “Organization and Reorganization of Immunoglo-
`3Llll1'1 Genes in A-MuLV-Transformed Cells: Rearrange-
`nent of Heavy but Not Light Chain Genes” Cell 272381-390
`(Dec. 1981).
`Altenburger et al., “Functional and non-functional joining in
`'mmunoglobulin light chain genes of a mouse myeloma”
`Nature 2872603-607 (Oct. 16, 1980).
`Amzel and Poljak, “Three-dimensional structure of immu-
`ioglobulins” Ann. Rev. Biochem. 482961-997 (1979).
`Astaldi et al., “Increase of hybridoma formation by human
`ymphocytes after stimulation in vitro; effect of antigen,
`endothelial cells, and PWM” J. Immunol. 128(6)22539-2542
`(1982).
`August, “Monoclonal Antibodies—I2 Discussion” Cell
`Fusion: Gene Transfer and Transformation, Beers et al. pp.
`345-351 (1984).
`Barnett-Foster and Painter, “The interaction of the Facb
`fragment of rabbit anti-sheep red cell IgG With guinea pig
`macrophages, and human monocytes and granulocytes”
`lllolecular Immunology 19(2):247-252 (1982).
`Bernard and Gough, “Nucleotide sequence of immunoglo-
`bulin heavy chain joining segments between translocated VH
`and ‘u constant region genes” Proc. Natl. Acad. Sci. USA
`77(6)2363(L3634 (1980).
`Bernard et al., “Plasmacytomas with more than one immu-
`noglobulin K mRNA:
`Implications for allelic exclusion”
`Proc. Natl. Acad. Sci. USA 78(9)25812-5816 (Sep. 1981).
`Bernstein et al., “Monoclonal Antibody Therapy of Mouse
`Leukemia” Monoclonal Antibodies, Kennett et al., Plenum
`Press pp. 275-291 (1980).
`Better and Horwitz, “Expression of engineered antibodies
`and antibody fragments in microorganisms” Methods in
`Enzymology 1782476-496 (1989).
`Better et al., “Escherichia coli secretion of an active chi-
`meric antibody fragment” Science 24021041-1043 ( 1988).
`Bevan et al,. “Biosynthesis of immunoglobulins” Progress
`In Biophysics and Molecular Biology, Butler and Noble,
`Pergamon Press pp. 133-162 (1972).
`Birshtein et al., “Effects of immunoglobulin structure on Fc
`receptor bi11ding: a mouse myeloina variant in1n1unoglobu-
`lin with a y2b-y2a hybrid heavy chain having a complete y2a
`Fc region fails to bind a y2a Fc receptors on mouse mac-
`rophages” J. Immunol. 129(2)2610-614 (1982).
`Blythman et al., “Immunotoxins2 hybrid molecules of mon-
`clonal antibodies and a toxin subunit specifically kill tumour
`cells” Nature 2902145-146 (1981).
`Bobrzecka et al., “The method of controlled rearrangement
`of protein disulphides and its use for synthesis of chimeric
`immunoglobulin G” Immunology Letters 22151-155 (1980).
`Bock et al., “Cloning and expression of the cDNA for human
`antithrombin III” NucleicAcidsResearch 10(24)28113-8125
`(1 982).
`
`Genzyme Ex. 1001, pg 2
`
`Genzyme Ex. 1001, pg 2
`
`

`
`US 6,331,415 B1
`Page 3
`
`Bock et al., “Hybridization-selected translation of Bombyx
`mori high—cysteine chorion proteins in Xenopus laevis
`oocytes” Proc. Natl. Acad. Sci. USA 7921032-1036 (1982).
`Boss et al., “Assembly of functional antibodies from immu-
`noglobulin heavy and light chains synthesised in E. coli”
`Nucleic Acids Research 12(9)23791-3806 (1984).
`Boss et al., “Production of immunoglobulin molecules in
`Escherichia coli” J. Cell. Biochem. (Abstract Only) Supple-
`ment 7A:140 (0364) (1983).
`7» genes in the
`Bothwell et al., “Dual expression of
`MOPC-315 plasmacytoma” Nature 290265-67 (1981).
`Bothwell et al., “Heavy chain variable region contribution to
`the NP” family of antibodies: somatic mutation evident in a
`y2a variable region” Cell 242625-637 (1981).
`Bothwell et al., “Somatic variants of murine immunoglobu-
`lin (X light chains” Nature 2982380-382 (Jul. 22, 1982).
`Boulianne et al., “Production of functional chimaeric mouse/
`human antibody” Nature 312:643—646 (Dec. 13, 1984).
`Boulianne et al., “The production of chimeric mouse/human
`antibodies” Abstracts ofpapers presented at the meeting on
`Cellular and Molecular Biology of Neoplasia (Abstract
`only) pp. #25 (1983).
`Boyd et al., “Human monoclonal antibodies—production
`and potential” Trends in Biotechnology 2(3)270—77 (1984).
`Boyden, Alar1,
`“Homology
`and Analogy” Science
`164:455-456 (Apr. 1969).
`Boylston et al., “Production of human IgM anti—D in tissue
`culture by EB—virus—transformed lymphocytes” Scand. J.
`Immunol. 122355-358 (1980).
`Bruggemann et al., “Immunoglobulin V region variants in
`hybridoma cells. I. Isolation of a variant with altered idio-
`typic and antigen binding specificity” Ell/[B0 Journal
`1(5):629-634 (1982).
`Buchner et al., “Renaturation, purification and characteriza-
`tion of recombinant Fab-fragments produced in Escherichia
`coli” Bio/Technology 92157-162 (1991).
`Burrows et al., “Evidence that murine pre-B cells synthesize
`0 heavy chains but no light chains” Nature 2802838-841
`(Aug. 30, 1979).
`Cabilly et al., “Generation of Antibody Activity from Immu-
`noglobulin Polypeptide Chains Produced in Escherichia
`coli” Proc. Natl. Acad. Sci. USA 8123273—3277 (1984).
`Chang et al., “Construction and characterization of ampli-
`fiable multicopy DNA cloning vehicles derived from the
`P15A cryptic miniplasmid” J. Bacteriol. 134(3)21141—1156
`(1978).
`Cheng et al., “Effect of deglycosylation on the binding and
`immunoreactivity of human thyroxi11e—binding globulin”
`Journal of Biological Chemistry 254(18)28830-8835 (Sep.
`25, 1979).
`Choi et al., “RNA splicing generates a variant light chain
`from an aberrantly rearranged K gene” Nature 2862776-779
`(Aug. 21, 1980).
`Coffino and Laskov, “Immunoglobulin production: method
`for quantitatively detecting variant myeloma cells” Science
`1672186-188 (1970).
`Coffino et al., “Suppression of Immunoglobulin Synthesis
`by Cellular Hybridization” Nature New Biology 231287-90
`(May 19, 1971).
`Cook and Scharff, “Antigen-binding mutants of mouse
`myeloma
`cells”
`Proc.
`Natl. Acad.
`Sci.
`USA
`7(12):5687—5691 (1977).
`
`c11lt11red mouse
`Cook et al., “Somatic mutation in a
`myeloma cell affects antigen binding” Proc. Natl. Acad. Sci.
`USA 7921240-1244 (1982).
`Cotton and Milstein, “Fusion of two immunoglobulin-pro-
`ducing myeloma cells” Nature 24424243 (Jul. 6, 1973).
`Crews et al., “A Single VH Gene Segment Encodes the
`Immune Response to Phosphorylcholine2 Somatic Mutation
`Is Correlated with the Class of the Antibody” Cell 25:59-66
`(1981).
`Croce et al., “Production of human hybridomas secreting
`antibodies to measles virus” Nature 2882488-489 (1980).
`Dangl et al., “Segmental flexibility and complement fixation
`of genetically engineered chimeric human, rabbit and mouse
`antibodies” The EMBO Journal 7(7)21989-1994 (1988).
`Dangl, “Rapid isolation of cloned isotype switch variants
`using fluorescence
`activated cell
`sorting” Cytometry
`2(6):395-401 (1982).
`De Bernardez-Clark and Georgiou, “Inclusion bodies and
`recovery of proteins from the aggregated state” Protein
`Refolding Chapter 121-20 (1991).
`De Boer et al., “Construction of a Tandem trp-lac Promoter
`and a Hybrid trp-lac Promoter for Eflicient and Controlled
`Expression of the Human Growth hormone Gene in Escheri-
`chia coli " Promoters, Structure and Function (Praeger Pub-
`lishers, R. Rodriguez and M. Chamberline, eds.) pp.
`462-481 (1982).
`Dildrop et al., “Immunoglobulin V region variants in hybri-
`doma cells. II. Recombination between V genes” EMBO
`Journal 1(5)2635-640 (1982).
`Dover, Gabby, “Nonhomologous Views of a Terminology
`Muddle” Cell 512515-516 (Nov. 20, 1987).
`Duyvesteyn and De Waard, “A new sequence-specific endo-
`nuclease from a thermophilic cyanobacterium, mastigocla-
`dus laminosus” FEBS Letters 111(2)2423-426 (1980).
`Duyvesteyn et al., “Sequence-specific endonucleases in
`strains
`of
`anabaena
`and
`nostoc” Arch. Microbiol.
`
`1342276-281 (1983).
`Early and Hood, “Mouse immunoglobulin genes” Genetic
`Engineering, Principles and Methods, Setlow and H01-
`laender, N.Y. and London, UK2Plenum Press vol. 32157-188
`(1981).
`Early et al., “Allelic Exclusion and Nonproductive I111111u-
`noglobulin Gene Rearrangements” Cell 2421-3 (Apr. 1981).
`Early et al., “An immunoglobulin heavy chain variable
`region gene is generated from three segments of DNA: VH
`1) and 1,,” Cell 192981-992 (1980).
`Edelman, G., “Antibody structure and molecular immunol-
`ogy” Annals of the New YorkAcade/ny ofSciences 19025-25
`(1971).
`Edwards et al., “A human-human hybridoma system based
`on a fast—growing mutant of the ARH—77 plasma cell
`leukemia-derived line” European J. Immunol. 122641-648
`(1982).
`Ellison et al., “Linkage and sequence homology of two
`human immunoglobulin gamma heavy chain constant region
`genes” Proc. Natl. Acad. Sci. USA 7921984-1988 (Mar.
`1982).
`Ellison et al., “Nucleotide sequence of a human immuno-
`globulin CV4 gene” DNA 1(1):11-18 (1981).
`Ellison et al., “The nucleotide sequence of a human immu-
`noglobulin
`CV1
`gene” Nucleic Acids
`Research
`10(13):4071—4079 (1982).
`
`Genzyme Ex. 1001, pg 3
`
`Genzyme Ex. 1001, pg 3
`
`

`
`US 6,331,415 B1
`Page 4
`
`Eshhar et al., “Ind11ction of secretion of IgM from cells of
`the B cell line 38C-13 by somatic cell hybridization” J.
`Immunol. 122(6)22430-2434 (1979).
`Feiss et al., “Separate sites for binding and nicking of
`bacteriophage 7» DNA by terminase” Proc. Natl. Acad. Sci.
`USA 802955-959 (1983).
`Fiers et al., “Complete Nucleotide Sequence of SV40 DN ”
`Nature 2732113-120 (May 11, 1978).
`Gillies et al., “A tissue-specific transcription enhancer ele-
`ment is located in the major intron of a rearranged immu-
`noglobulin heavy chain gene” Cell 332717-728 (1983).
`Gillies et al., “Antigen Binding and Biological Activities of
`Engineered Mutant Chimeric Antibodies with Human
`Tumor Specificities” Hunt. Antibod. Hybridomas 1(1)247-54
`(1990).
`Givol et al., “Diversity of gerni-line inununoglobulin VH
`genes” Nature 2922426-430 (1981).
`Gluzman, Yakov, “SV40-Transformed Simian Cells Sup-
`port
`the Replication of Early SV40 Mutants” Cell
`232175-182 (Jan. 1981).
`Goldsby et al., “Hybrid cell lines with T—cell characteris-
`tics” Nature 2672707-708 (Jun. 23, 1977).
`Graham and van der Eb, “A New Technique for the Assay of
`Infectivity of Human Adenovirus 5 DN ” Virology
`522456-467 (1973).
`Gritz and Davies, “Plasmid-encoded hygromycin B resis-
`tance:
`the sequence of hygromycin B phosphotransferase
`gene and its expression in Escherichia coli and Saccharo-
`myces cerevisiae” Gene 252179-188 (1983).
`Gupta et al., “General Orientation of Human Lymphocyte
`Subpopulations” Clinical Immunobiol, Bach and Good,
`Academic Press vol. 4:1-18 (1980).
`Hedin et al., “Specificities and Binding Properties of Eight
`Monoclonal Antibodies Against Carcinoembryonic Anti-
`gen” Molecular Immunology 1921641-1648 (1982).
`Herlyn et al., “Inhibition of Growth of Colorectal Carcinoma
`in Nude Mice by Monoclonal Antibody” Cancer Research
`402717-721 (Mar. 1980).
`Herzenberg et al., “Hybridoma Variants Affecting Isotype,
`Antigen Binding, and Idiotype” Biotechnology in Diagnos-
`tics, Koprowski et al. vol. 2123-16 (1985).
`Hieter et al., “Cloned human and mouse kappa immunoglo-
`bulin constant and J region genes conserve homology in
`functional segments” Cell 22(Part 1)2197-207 (1980).
`Honjo et al., “Cloning and complete nucleotide sequence of
`mouse immunoglobulin y1 Chain Gene” Cell 182559-568
`(1979).
`Honjo et al., “Rearrangenients of Inununoglobulin Genes
`during Differentiation and Evolution” Immunological Rev.
`59:33-67 (1981).
`Hood et al., “Antibodies” Immunology, Forkner and Moore,
`Philippines2The Benjamin/Cummings Publishing Co., Inc.,
`Chapter 3, pp. 199-221 (1978).
`Ilorwitz et al., “Secretion of functional antibody and Fab
`fragment from yeast cells” Proc. Natl. Acad. Sci. USA
`8528678—8682 (1988).
`Houghton ct al., “Detection of cell surface and intracellular
`antigens by human monoclonal antibodies” Journal of
`Experimental Medicine 158253-65 (1983).
`Howard et al., “A Rapid Method for the Detection of
`Antibodies to Cell Surface Antigens: A Solid Phase Radio-
`immunoassay Using Cell Membranes” Journal of Immuno-
`logical Methods 38:75-84 (1980).
`
`Howard et al., “Long-term culture of normal mouse B
`lymphocytes” Proc. Natl. Acad. Sci. USA 78(9)25788-5792
`(Sep. 1981).
`Hughes and Murray, “The nucleotide sequences recognized
`by endonucleases Aval and AvaII from Anabaena variabi-
`lis“ Biochemical Journal 185265-75 (1980).
`Hunkapiller et al., “The growing immunoglobulin gene
`superfamily” Nature 323215-16 (1986).
`“Immunoglobulin molecules
`and genes” Microbiology
`Including Immunology and Molecular Genetics, Third edi-
`tion, Harper International Edition vol. Chapter 172338-379.
`Inouye et al., “Signal Sequence of Alkaline Phosphatase of
`Escherichia coli” J. Bacteriol. 1492434 (1982).
`Isenman et al., “The structure and function of immunoglo-
`bulin domains” J. Immunol. 114(6)21726-1929 (1975).
`Itakura and Riggs, “Chemical DNA synthesis and recombi-
`nant DNA studies” Science 20921401-1405 (1980).
`Jaton et al., “Conformational changes induced in a homo-
`geneous anti-type III pneumococcal antibody by oligosac-
`charides of increasing size” Biochemistry 14(24)25312-5315
`(1975).
`Johnson et al., “The Complete V Domain Amino Acid
`Sequences of Two Myeloma Inulin-Binding Proteins” Jour-
`nal of Immunology 1282302-307 (1982).
`Jones, P.T. et al., “Replacing the Complementarity-deter-
`mining Regions in a Human Antibody with Those From a
`Mouse” Nature 3212522-525 (May 29, 1986).
`Kabat, E., “Activation of the complement system and its
`effect on cells” Structural Concepts in Inzmunology and
`Immunochemistry, Second edition, IIolt, Rinehart and Win-
`ston vol. Chapter 132413-435 (1976).
`Kaivarainen et al., “Hapten-induced changes in pig anti-
`-Dansyl antibodies revealed by EPR spectra of spin-la-
`belled antibodies” Immunol. Letters 325-11 (1981).
`Kalderon et al., “Deletion loop mutagenesisz a novel method
`for
`the construction of point mutations using deletion
`mutants” Nucl. Acids Res. 1025161-5168 (1982).
`Keshet et al, “Cloning of bovine growth hormone gene and
`its expression in bacteria” Nucleic Acids Research 9:19-30
`(1981).
`Kipps et al., “Allotype Switch Variants in Cultured Mono-
`clonal Producing Hybridomas” Journal of Cellular Bio-
`chemistry (abstract only) pp. 163 (-1984).
`Kitai et al., “Extracellular production of human immuno-
`globulin G FC region” Microbiol. Biotechnol. 28(1):52-56
`(1988).
`Klein et al., “Expression of biological effector functions by
`immunoglobulin G molecules lacking the hinge region”
`Proc. Natl. Acad. Sci. USA 78(1)2524-528 (1981).
`Kohl and Moore, “Human antibody-dependent cellular
`cytotoxicity and natural killer cytotoxicity to herpes simplex
`virus-infected autologous and allogeneic cells” Immunology
`482187-193 (1983).
`Kohler et al., “Derivation of Specific Antibody-producing
`Tissue Culture and Tumor Lines by Cell Fusion” European
`Journal ofImmunology 62511-519 (1976).
`Kohler, G. and Milstein, C., “Continuous Cultures of Fused
`Cells Secreting Antibody of Predefined Specificity” Nature
`256:495—497 (Aug. 7, 1975).
`Konieczny et al., “The combination of IgM subunits and
`proteolytic IgG fragments by controlled formation of inter-
`chain disulphides” Haematologia 14(1)295-99 (1981).
`
`Genzyme Ex. 1001, pg 4
`
`Genzyme Ex. 1001, pg 4
`
`

`
`US 6,331,415 B1
`Page 5
`
`Koskimies, S., “Human lymphoblastoid cell line producing
`specific antibody against Rh-antigen D” Scand. J. Immunol.
`11:73-77 (1980).
`Kuehl, W.M., “Light chain fragments: aberrant expression of
`immunoglobulin genes” TIBS pp. 206-208 (Aug. 1981).
`Kupchik et al., “Monoclonal Antibodies to Carcinoembry-
`onic Antigen Produced by Somatic Cell Fusion” Cancer
`Research 41:3306-3310 (Sep. 1981).
`Kwan, “Two Kappa Immunoglobulin Genes Are Expressed
`in the Myeloma S107” Cell 26:57-66 (Oct. 1981).
`Larson et al., “Saccharomyces cerevisiae actin-Escherichia
`coli lacZ gene fusions: synthetic-oligonucleotide-mediated
`deletion of the 309 base pair intervening sequence in the
`actin gene” Gene 22:31-39 (1983).
`Laskov and Scharff, “Synthesis, assembly, and secretion of
`gamma globulin by mouse myeloma cells” Journal of
`Experimental Medicine 131(3):515-541 (1970).
`Laskov et al., “Induction of amplified synthesis and secre-
`tion of IgM by fusion of murine B Lymphoma with myeloma
`cells” Proc. Natl. Acad. Sci. USA 76(2):915-919 (Feb.
`1979).
`Lau and Doolittle, “Aqu I: a more easily purified isoschi-
`zomer of AVA I” FEBS Letters 121(2):200-202 (1980).
`Leder, P., “The genetics of antibody diversity” Scientific
`America 246:72-83 (1982).
`Lee et al., “Characterization of the Gene Encoding Heat-
`Stable Toxin II and Preliminary Molecule Epidemiological
`Studies of Enterotoxigenic Escherichia coli Heat—Stable
`Toxin II Producers” Infection and Immunity 42:264-268
`(Oct. 1983).
`Letherbarrow et al., “Effector functions of a monoclonal
`aglycosylated mouse IgGza:binding and activation of
`complement component C1 and interaction with human
`monocyte Fc receptor” Molec. Immunol. 22(4):407-415
`(1985).
`Levy and Miller, “Tumor therapy with monoclonal antibod-
`ies” Fed. Proc. 42:2650-2656 (1983).
`Lewin, Roger, “When Does Homology Mean Something
`Else?” Science 23721570 (1987).
`Liu et al., “Chimeric mouse-human IgG1 antibody that can
`mediate lysis of cancer cells” Proc. Natl. Acad. Sci. USA
`84:3439—3443 (May 1987).
`Liu, Pinghui V., “Pseudomonas Toxins” J.
`130:S94-S99 (1974).
`Vlaniatis et al. Molecular Cloning: A Laboratory ll/Ianual,
`1st edition, New York: Cold Spring Harbor Lab Press, Chap-
`ter 12, pps. selected pages (1982).
`Vlaniatis, T., “Recombinant DNA procedures in the study of
`eukaryotic genes” Cell Biol. 3:563-608 (1980).
`Vlargu lies et al., “Regulation of imm11noglob11 lin expression
`in mouse myeloma cells” Immunoglobulin Expression pp.
`781-791 (1977).
`Vlargulies et al., “Somatic Cell Hybridization of Mouse
`Vlycloma” Cell 8:405-415 (Jul. 1976).
`'VIartinis et al., “Monoclonal antibodies with dual antigen
`specificity” Oncology pp. 311-316.
`Vlather et al., “Transcriptional regulation of immunoglobu-
`lin V genes” Nucleic Acids Research 9(24):6855-6867
`(1981).
`Vlatsuuchi and Morrison, “Antigen binding variants of
`mouse plasmacytoma J558” Fed. Proc.
`(Abstract only)
`37:1763 (2703) (1978).
`
`Infect. Dis.
`
`Max et al., “Variation in the Crossover Point of Kappa
`Immunoglobulin Gene V-J Recombination: Evidence from
`a Cryptic Gene” Cell 21:793-799 (Oct. 1980).
`\/IcCutchan et al., “Enhancement of the Infectivity of Simian
`Virus 40 Deoxyribonucleic Acid with Diethylaminoethyl-
`Dextran” J. Natl. Cancer Institute 41:351-356 (1968).
`\/Ielchers, F., “Biosynthesis of the carbohydrate portion of
`'mmunoglobulin radiochemical and chemical analysis of the
`carbohydrate moieties of two myeloma proteins purified
`rom different subcellular fractions of plasma cells” Bio-
`chemistry 10(4):653-659 (1971).
`\/Iertz et al., “Defective Simian Vir11s 40 Genomes: Isolation
`and Growth of Individual Clones” Wrology 62:112-124
`(1974).
`\/Iessing et al., “A System for Shotgun DNA Sequencing”
`Nucleic Acids Research 9(2):309-321 (1981).
`\/Iilstein et al., “Clonal Variants of Myeloma Cells” Progress
`in Immunology II 1:157-168 (1974).
`\/Iohit and Fan, “Hybrid Cell Line from a Cloned Immuno-
`globulin-Producing Mouse Myeloma and a Nonproducing
`\/Iouse Lymphoma” Science 171175-77 (Jan. 8, 1971).
`\/Iorrison and Scharff,
`“Mutational events
`in mouse
`myeloma cells” Critical Reviews in Immunology 3(1):1-22
`(1981).
`\/Iorrison et al., “Chimeric Human Antibody Molecules:
`\/Iouse Antigen-binding Domains with Human Constant
`Region Domains”Proc. Natl.Acad. Sci. USA 81:6851-6855
`(Nov. 1984).
`\/Iorrison et al., “Genetically Engineered Antibody Mol-
`ecules” Adv Immunol. 44:65-92 (1989).
`\/Iorrison et al., “Transfectomas Provide Antibodies With
`\Iovel Structures and Functions”Antibodies.' Structure, Syn-
`thesis, Function, and Immunologic Intervention in Disease,
`Szentivanyi et al. pp. 167-178 (1987).
`\/Iorrison et al., “Transfer and expression of immunoglobu-
`in genes”Annual Review ofImmunologv 2:239-256 (1984).
`\/Iorrison, S. L., “Transfectomas Provide Novel Chimeric
`Antibodies” Science 229:1202-1207 (Sep. 20, 1985).
`\/Iovva et al., “Amino Acid Sequence of the Signal Peptide
`of ompA Protein, a Major Outer Membrane Protein of
`Escherichia coli” The Journal of Biological Chemistry
`255:27-29 (Jan. 10, 1980).
`\Ial<abayashi et al., “The transforming function of bovine
`aapillomavirus DNA” Proc. Natl. Acad.
`Sci. USA
`80:5832-5836 (1983).
`\Ieuberger et al., “A hapten-specific chimaeric IgE antibody
`with human physiological
`effector
`function” Nature
`3142268-270 (Mar. 21, 1985).
`\Ieuberger et al., “Recombinant Antibodies Possessing
`\Iovel Effector Functions” Nature 312:604-608 (Dec. 13,
`1984).
`\Ieuberger, “Switch from hapten-specific immunoglobulin
`VI to immunoglobulin D secretion in a hybrid mouse cell
`ine” Proc. Natl. Acad. Sci. USA 78(2):1138-1142 (1981).
`Ochi et al., “Functional immunoglobulin M production after
`ransfection of cloned immunoglobulin heavy and light
`chain genes into lymphoid cells” Proc. Natl. Acad. Sci. USA
`80:6351-6355 (1983).
`Oi et al., “Correlation between segmental flexibility and
`effector function of antibodies” Nature 307:136-140 (1984).
`Oi et al., “Hybridoma Antibody-Producing Switch Variants:
`A Variant Lacking the CH1 Domain” Cell Fusion.‘ Gene
`Transfer and Transformation, R.F. Beers, Jr. and E.G. Bas-
`sett, Raven Press, New York pp. 281-287 (1984).
`
`Genzyme Ex. 1001, pg 5
`
`Genzyme Ex. 1001, pg 5
`
`

`
`US 6,331,415 B1
`Page 6
`
`Oi et al., “Localization of M11rine Igh-1“ Allotypic Deter-
`minants by Using a Panel of Mouse Myeloma Variant
`Immunoglobulins”
`Journal
`of
`Immunology
`130(4)21967-1969 (Apr. 1983).
`Oi et al., “Lymphocyte membrane IgG and secreted IgG are
`structurally ar1d allotypically distinct” Journal of Experi-
`mental Medicine 15121260-1274 (1980).
`Olsson and Kaplan, “Human-human hybridomas producing
`monoclonal antibodies of predefined antigenic specificity”
`Proc. Natl. Acad. Sci. USA 77(9)25429-5431 (1980).
`Orna Zemel-Dreasen et al., “Secretion and processing of an
`immunoglobulin light chain in Escherichia coli” Gene
`27(3):315-322 (1984).
`Owens and Young, “The genetic engineering of monoclonal
`antibodies” J. Immunol. It/[ethods 168:149-165 (1994).
`Palva et al., “Secretion of interferon by bacillus subtilis”
`Gene 222229-235 (1983).
`Parham et al., “Isolation of Heavy Chain Class Switch
`Variants of a Monoclonal Anti-DC1 Hybridoma Cell Line:
`Effective Conversion of Noncytotoxic IgG1 Antibodies to
`Cytotoxic IgG2 Antibodies” Human Immunology 82141-151
`(1983).
`Peden and Nathans, “Local mutagenesis within deletion
`loops of DNA heteroduplexes” Proc. Natl. Acad. Sci. USA
`79:7214—7217 (1982).
`Periman, “IgG Synthesis in Hybrid Cells from an Anti-
`body-producing Mouse Myeloma and an L Cell Substrain”
`Nature 228:1086—1087 (Dec. 12, 1970).
`Perry et al., “Transcription of r11ouse K chain ger1es: irnpli-
`cations for allelic exclusion” Proc. Natl. Acad. Sci. USA
`77(4):1937—1941 (1980).
`Picken et al., “Nucleotide sequence of the gene for heat-
`-stable enterotoxin II of Escherichia coli” Infection and
`Immunity 42(1)2269-275 (1983).
`Plaut et al., “Immunoglobulin M: Fixation of human
`complement by the Fc Fragment” Science 176255-56
`(1972).
`Ponte et al., “Transcription of immunoglobulin heavy-chain
`sequences from the excluded allele” Nature 2912594-596
`(1981).
`Raghunathan et al., “Abstract P-D2-01-Influence of vari-
`able domain glycosylation on antigen binding2crystal struc-
`ture of anti-GnRH Fab frament” Prog. Biophy & Mol. Biol.
`65(5):143—(1996).
`Raschke et al., “Assembly and secretion of pentameric IgM
`in a fusion between a nonsecreting B cell lymphoma and an
`IgG-secreting plasmacytoma” Proc. Natl. Acad. Sci. USA
`76(7):3469-3473 (1979).
`Rechavi et al., “Evolutionary aspects of immunoglobulin
`heavy chain variable region (VI, gene subgroups” Proc.
`Natl. Acad. Sci. USA 80:855-859 (1983).
`Reeck et al., “Homology” in Proteins and Nucleic Acids: A
`Terminology Muddle and a Way out of it” Cell 50 (Aug. 28,
`1987).
`Reth et al., “Analysis of the repertoire of anti-NP antibodies
`in C57BL/6 mice by cell fusion” European Journal of
`Immunology 82393-400 (1978).
`Rice et al, “Measurement of transient cDNA expression in
`mammalian cells using flow cytometric cell analysis and
`sorting” Cytometry 122221-233 (1991).
`Riley et al., “Induction of light chain expression in a pre-B
`cell line by fusion to myeloma cells” Nature 2892804-806
`(1981).
`
`Robins et al., “Regulated Expression of Human Growth
`Hormone Genes in Mouse Cells” Cell 292623-631 (1982).
`Roizes, “A new specific endonuclease from Anabaena vari-
`abilis” FEBS Letters 104(1):39-44 (1979).
`Rouger and Goossens, “Human Monoclonal antibodies
`against human red blood cells” Therapeutic Monoclonal
`Antibodies, Borrebaeck and Larrick, M Stockton Press pp.
`263-286 (1990).
`Rybarska et al., “The Hemolytic Activity of (Fab-Fc)
`Recombinant
`Immunoglobulins with Specificity for the
`Sheep Red Blood Cells” Immunology Letters 42279-284
`(1982).
`Sakano et al., “Domains and the hinge region of an immu-
`noglobulin heavy chain are encoded in separate DNA seg-
`ments” Nature 2772627-633 (1979).
`Scharff et al., “Variations ir1 the synthesis and assembly of
`immunoglobulins by mouse myeloma cells: A Genetic and
`Biochemical Analysis” J. Cellular Physiology 762331-348
`(1970).
`Scholm et al., “Generation of human monoclonal antibodies
`reactive with human mammary carcinoma cells” Proc. Natl.
`Acad. Sci. USA 77(11)26841—6845 (1980).
`Schroder et al., “Florescence-activated cell sorting of
`mouse-human hybrid cells aids in locating the gene for the
`Leu 7 (HNK-1) antigen to human chromosome 11” Proc.
`Natl. Acad. Sci. USA 8023421-3424 (Jun. 1983).
`Schwaber and Cohen, “Human x mouse somatic cell hybrid
`clone secreting immunoglobulins of both parental types”
`Nature 2442444