aNmeloTt
`
`Biochemistry, Genetics,
`um(Teh(aelweNa)aesLh)kS
`
`THEODORE PETERS,JR.
`
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`

`

`All about Albumin
`Biochemistry, Genetics, and Medical Applications
`
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`A medic administering blood plasma (shortly to be replaced by purified albu-
`min) to a wounded U.S. Soldier in Sicily in 1943. With appreciation to the
`National Library of Medicine.
`
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`

`

`All about Albumin
`Biochemistry, Genetics, and Medical Applications
`
`By Theodore Peters, Jr.
`Research Institute
`The Mary Imogene Bassett Hospital
`Cooperstown, New York
`
`ACADEMIC PRESS
`A n I m p r i n t o f Elsevier
`
`San Diego New York Boston London Sydney Tokyo Toronto
`
`MPI EXHIBIT 1042 PAGE 5
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`

`

`Front cover photograph: Ribbon diagram of tertiary structure
`of human albumin as derived by X-ray crystallography. Reproduced
`from He and Carter (1992) by permission of Nature.
`
`This book is printed on acid-free paper. ( ~
`
`Copyright (cid:14)9 1996 by ACADEMIC PRESS
`
`All Rights Reserved.
`No part of this publication may be reproduced or transmitted in any form or by any
`means, electronic or mechanical, including photocopy, recording, or any information
`storage and retrieval system, without permission in writing from the publisher.
`
`Permissions may be sought directly from Elsevier's Science and Technology Rights Department in
`Oxford, UK. Phone: (44) 1865 843830, Fax: (44) 1865 853333, e-mail: permissions@elsevier.co.uk.
`You may also complete your request on-line via the Elsevier homepage: http:Hwww.elsevier.com by
`selecting "Customer Support" and then "Obtaining Permissions".
`
`A c a d e m i c P r e s s
`An Imprint of Elsevier
`525 B Street, Suite 1900, San Diego, California 92101-4495, USA
`http://www.apnet.com
`
`A c a d e m i c P r e s s
`Harcourt Place, 32 Jamestown Road, London NWI 7BY, UK
`http://www.hbuk.co.uk/ap/
`
`Library of Congress Cataloging-in-Publication Data
`
`Peters, Theodore, date.
`All about albumin : biochemistry, genetics, and medical
`applications / by Theodore Peters, Jr.
`p.
`em.
`Includes bibliographical references and index.
`ISBN 0-12-552110-3 (alk. paper)
`1. Serum albumin.
`I. Title.
`QP99.3.A4P48
`1995
`574.19 . . . . . . . .
`/ ~ . . ) l . - - u C L u
`
`CIP
`
`PRINTED IN THE UNITED STATES OF A M E R I C A
`05 SB 11 10 9 8 7 6 5 4
`
`('1~ ")1 1 A ' I
`7J-~.. 1 1 '~J
`
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`

`

`To the thousands of students of albumin who have
`
`contributed to our knowledge of this captivating
`
`protein over many years. And particularly to my
`
`lovely wife and companion of over 50 years,
`Margaret Campbell Peters, who has supported
`
`my efforts and tolerated this rival without jealousy.
`
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`

`

`Contents
`
`Foreword: A l l about A l b u m i n
`
`.......................................................................
`
`Preface
`
`...........................................................................................................
`
`xi
`
`xv
`
`List of Abbreviations
`
`.....................................................................................
`
`xix
`
`1 H i s t o r i c a l P e r s p e c t i v e
`
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`1
`
`2 The Albumin Molecule" Its Structure and Chemical Properties
`I.
`P r i m a r y S t r u c t u r e a n d C o m p o s i t i o n
`...............................................................
`
`A. A m i n o Acid Sequence and Disulfide Bonding Patterns
`
`..........................................
`
`9
`
`9
`
`B. Composition
`
`............................................................................................................ 14
`
`C. Features
`
`................................................................................................................... 17
`
`D. Fragments
`
`................................................................................................................ 19
`
`II.
`
`T e r t i a r y S t r u c t u r e a n d P h y s i c a l C h e m i c a l B e h a v i o r .......................................... 24
`
`A. Configuration
`
`.......................................................................................................... 24
`
`B. Physical Chemistry
`
`................................................................................................. 39
`
`C. Changes in Configuration
`
`....................................................................................... 54
`
`3 Ligand Binding by Albumin
`I. A n i o n i c a n d N e u t r a l L i g a n d s
`........................................................................... 79
`
`A. Long-Chain Fatty Acids
`
`........................................................................................... 79
`
`B. Bilirubin
`
`.................................................................................................................. 95
`
`o
`
`C. Site-I Ligands
`
`.......................................................................................................... 102
`
`D. Site-II Ligands
`
`........................................................................................................ 109
`
`E. Miscellaneous Anionic and Neutral Ligands
`
`.......................................................... 116
`
`II.
`
`C a t i o n i c L i g a n d s
`
`............................................................................................. 121
`
`A. Copper(II) and Nickel(II)
`
`........................................................................................ 121
`
`vii
`
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`

`

`viii
`
`Contents
`
`B. Calcium and Magnesium
`
`........................................................................................ 124
`
`C. Other Metals and Cationic Drugs
`
`............................................................................ 126
`
`III.
`
`A n t i b o d i e s ; I m m u n o c h e m i s t r y o f A l b u m i n
`
`.................................................... 127
`
`A. Number and Distribution of Determinants
`
`.............................................................. 128
`
`B. Sequences and Chemical Groups Involved
`
`............................................................. 129
`
`C. Albumin Immunology in V i v o
`
`................................................................................. 131
`
`4 Genetics: The Albumin Gene
`I. G e n e S t r u c t u r e
`................................................................................................ 133
`
`A. Albumin Gene Sequence
`
`...................................................................... . .................. 134
`
`B. Codon Usage
`
`........................................................................................................... 141
`
`C. Polymorphisms
`
`....................................................................................................... 142
`
`II.
`
`C l o s e R e l a t i v e s : T h e A l b u m i n S u p e r f a m i l y
`
`................................................... 143
`
`A. ot-Fetoprotein
`
`.......................................................................................................... 143
`
`B. or-Albumin
`
`............................................................................................................... 149
`
`C. Vitamin D-Binding Protein
`
`..................................................................................... 150
`
`III.
`
`E v o l u t i o n : O r i g i n s in the Past
`
`......................................................................... 153
`
`A.
`
`Identification
`
`........................................................................................................... 154
`
`B. Possible Origins
`
`...................................................................................................... 167
`
`IV. M u t a n t F o r m s
`
`.................................................................................................. 170
`
`A. Classification
`
`B. Molecular Location of Mutations
`
`........................................................................................................... 170
`........................................................................... 171
`
`C. Distribution in Populations
`
`..................................................................................... 173
`
`D. Effects of Mutations on Albumin Molecule
`............................................................ 178
`.................................................................................... 181
`
`E. Genetics of Analbuminemia
`
`5 Metabolism: Albumin in the Body
`I.
`B i o s y n t h e s i s
`..................................................................................................... 188
`
`A. Systems Used to Study Albumin Biosynthesis
`
`....................................................... 189
`
`B. Expression (Transcription)
`
`...................................................................................... 192
`
`C. Translation
`
`............................................................................................................... 206
`
`D. Secretion: Proalbumin and Its Possible Roles
`
`........................................................ 213
`
`E. Rate of Biosynthesis
`
`............................................................................................... 223
`
`II. D i s t r i b u t i o n , F u n c t i o n s , a n d F a t e in the B o d y
`
`................................................ 228
`
`A. Distribution
`
`............................................................................................................. 228
`
`B. Functions
`
`................................................................................................................. 234
`
`C. Survival i n A n a l b u m i n e m i a
`
`.................................................................................... 240
`
`D. Changes to Albumin in Circulation
`
`......................................................................... 243
`
`III. D e g r a d a t i o n : R o l e in N u t r i t i o n
`
`....................................................................... 245
`
`A. Rate of Degradation
`
`................................................................................................ 245
`
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`

`

`Contents
`
`ix
`
`B. Site(s) of Degradation
`
`............................................................................................. 247
`
`C. Mechanism of Degradation
`
`..................................................................................... 248
`
`D. Fate of Degradation Products
`
`.................................................................................. 249
`
`6 Clinical Aspects" Albumin in Medicine
`I. A s s a y s
`............................................................................................................. 251
`
`A. Methods
`
`................................................................................................................... 251
`
`B. Standardization
`
`....................................................................................................... 255
`
`II.
`
`P a t h o l o g y : C h a n g e s in D i s e a s e
`
`....................................................................... 2 5 6
`
`A. Albumin Concentration and Its Significance
`
`.......................................................... 256
`
`B. Relation of Albumin to Some Metabolic Diseases
`
`................................................. 260
`
`III.
`
`P a r e n t e r a l U s e s
`
`............................................................................................... 2 7 8
`
`A. Circulatory Support
`
`................................................................................................. 279
`
`B. Digestive Support ..................................................................................................... 280
`
`C. Removal of Toxins
`
`.................................................................................................. 281
`
`D.
`
`Imaging
`
`................................................................................................................... 281
`
`E. Drug Delivery
`
`......................................................................................................... 283
`
`E Coating Surfaces
`
`..................................................................................................... 284
`
`7 Practical Aspects" Albumin in the Laboratory
`I. M e t h o d s o f P r e p a r a t i o n
`................................................................................... 2 8 5
`
`A. Laboratory Purification
`
`........................................................................................... 286
`
`B. Commercial Purification
`
`......................................................................................... 291
`
`C. Recombinant Production
`
`......................................................................................... 296
`
`II.
`
`F u r t h e r P u r i f i c a t i o n : A l b u m i n H e t e r o g e n e i t y
`
`................................................ 2 9 8
`
`A. Associated Substances
`
`............................................................................................ 298
`
`B. Forms of Albumin Observed
`
`................................................................................... 301
`
`III.
`
`A l b u m i n P r o d u c t s a n d T h e i r in Vitro A p p l i c a t i o n s
`
`......................................... 3 0 5
`
`A. Products
`................................................................................................................... 305
`B.
`In Vitro Applications
`
`............................................................................................... 308
`
`IV.
`
`T e s t e d P r o c e d u r e s for U s e in L a b o r a t o r y
`
`....................................................... 3 1 2
`
`A. Preparing Solutions of Albumin
`
`............................................................................... 313
`
`B. Characterizing Albumin Preparations
`
`..................................................................... 313
`
`C.
`
`Isolating Albumin from Serum
`
`............................................................................... 315
`
`D. Modifying Albumin
`
`................................................................................................ 316
`
`.................................................................................................. 3 1 9
`B i b l i o g r a p h y
`I n d e x
`.............................................................................................................. 4 1 5
`
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`

`

`Foreword" All about Albumin
`
`Albumin is one of the longest known and probably the most studied of all
`proteins. Its manifold diverse functions have attracted the interest of scientists
`and physicians for generations. Its applications are many, both in clinical medi-
`cine and in basic research. Yet, until now, no monograph has been published that
`brings together the full scope of albumin: its history, structure, physical and bio-
`logical properties, genetics, metabolism, clinical applications, and its prepara-
`tions and uses in the laboratory. This book is timely, and no one is more quali-
`fied to write it than Dr. Peters. He has spent a lifetime in the study of albumin,
`and has also been the colleague, advisor, and friend of many whose research has
`contributed to knowledge of this protein.
`Albumin is the most abundant soluble protein in the body of all vertebrates
`and is the most prominent protein in plasma. Some of its physiological proper-
`ties have been recognized since the time of Hippocrates; albumin was named
`and first studied a century and half ago and was crystallized a century ago. Yet,
`the recent elucidation of its three-dimensional structure depended on crystalliza-
`tion in the space shuttle and recombinant technology. The physiological func-
`tions of albumin were the prime incentive for the intensive wartime program of
`plasma fractionation beginning in 1942 at Harvard. Not only did this program,
`and its commercial and university affiliates, produce tons of highly purified, sta-
`ble, virus-free albumin for battlefield use, it also provided the methodology for
`the purification of many other plasma proteins. In peacetime this led to a
`national program for blood procurement and plasma fractionation and the devel-
`opment of other products for clinical use such as gamma globulin and clotting
`factors. The availability of so much pure albumin, at a time when other proteins
`had to be purified laboriously, made aIbumin the favorite model for study by
`protein chemists. This prompted a voluminous increase in literature that has not
`abated to this day. Only a person with a lifetime of devotion to albumin could
`put this vast literature into perspective and summarize and interpret it as
`Dr. Peters has done.
`
`xi
`
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`

`

`xii
`
`Foreword
`
`The structure, properties, and ligand binding of albumin, described in Chap-
`ters 2 and 3, are intimately connected and constitute the prime interest of protein
`chemists, biochemists, and pharmacologists. The repeating pattern of three
`largely helical domains stabilized by multiple disulfide bonds is unique to the
`albumin family of proteins. Albumin itself is unique in its myriad affinities. How
`a protein with only two major binding sites can exhibit such diverse affinities is a
`puzzle to the protein chemist and a predicament for the pharmacologist. Crystal-
`lographic study of the binding of ligands has elucidated the complex nature of
`the sites in some instances, but many cases remain to be explained.
`Unlike many other plasma proteins that exhibit polymorphisms and muta-
`tions, some of which are harmful, geneti c variants of human albumin are rare
`and benign; hence, until recently there was little study of albumin mutations.
`However, after the protein sequence and, later, the gene and genomic sequences
`discussed in Chapter 4 were determined, identification of the cause of genetic
`variations of albumin was undertaken. More than 50 mutations have been identi-
`fied: point mutants, frameshifts, and splicing errors. None of these is pathologi-
`cal, but analbuminemia is paradoxical. How can the very rare persons who
`essentially lack albumin exhibit only minor symptoms when it has been shown
`that such important properties as maintenance of blood volume and binding and
`transport of metabolites and drugs are prototypical properties of albumin? The
`albumin superfamily, initially comprising albumin, a-fetoprotein, and vitamin
`D-binding protein, has recently been augmented by the discovery of afamin, or
`a-albumin. Chapter 4 shows that the structural relationships within this family
`and the sequence homology of albumins of species ranging from man to the lam-
`prey are providing insights into the evolution of albumin.
`The metabolism and clinical aspects of albumin are interrelated but are
`described separately in Chapters 5 and 6, respectively. As a nonglycosylated single
`chain of 585 amino acids tightly folded into three domains by 17 intrachain disul-
`fide bonds, albumin offers a revealing model for study of the processes of expres-
`sion, secretion, and intramolecular folding to produce the mature protein. Prompted
`by its multiple functions, many investigators have analyzed the mechanism and rate
`of biosynthesis and the catabolism of albumin, as well as its distribution in the
`serous fluids of the body. Changes in serum albumin concentration in disease, typi-
`cally the marked decline in malnutrition and in renal and liver disease, have long
`served as diagnostic and prognostic criteria. Because of potential viral contamina-
`tion of blood and plasma, notably hepatitis and AIDS, albumin, which is readily
`rendered virus-free by heating for 10 hours at 60~
`has been widely used in
`surgery and in the treatment of shock and trauma. It is this application, approaching
`100,000 kg a year in North America alone, that has required large-scale methods of
`commercial production and has recently prompted the application of recombinant
`technology. These methods are described in Chapter 7, which also illustrates the
`many in vitro applications familiar to the biochemist and microbiologist.
`
`MPI EXHIBIT 1042 PAGE 14
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`

`

`Foreword
`
`xiii
`
`tells it all. In the many areas described
`My title "All about Albumin"*
`above, this long-needed monograph will serve as a handbook for the experienced
`investigator and as an invaluable reference source for all who have a need to
`know about albumin. It deserves a place on the shelves of all libraries of medi-
`cine and basic medical sciences, and of biology and chemistry. I r e c o m m e n d it
`highly to physicians, clinical investigators, biochemists, protein chemists, phar-
`macologists, biologists, and chemists.
`
`Frank W. Putnam
`Indiana University
`Bloomington, Indiana
`
`* Author's Note: The publisher and I found Dr. Putnam's title an appealing one and so adopted
`it, somewhat presumptuously, as the title for the volume. Our thanks to Dr. Putnam.
`
`MPI EXHIBIT 1042 PAGE 15
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`MPI EXHIBIT 1042 PAGE 16
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`

`

`Preface
`
`Every student of biology and medicine is familiar with serum albumin as
`the most abundant and most easily measured protein of blood plasma. One of the
`purest proteins available commercially at a reasonable cost, albumin is also
`known to clinicians, nutritionists, physical chemists, biochemists, immunolo-
`gists, and, more recently, to geneticists and molecular biologists. Medically, a
`generous concentration of albumin in the bloodstream is a measure of the "Qual-
`ity of Life" (Kobayashi et al., 1991). The value of its commercial production
`exceeds one billion dollars annually. Yet there have been few reviews of the
`chemistry of albumin, and no monographs on this protein other than reports
`from two conferences. Hence, on retiring from laboratory activity after a lifetime
`of interest in this intriguing protein, I undertook the somewhat ambitious task of
`summarizing in one volume its chemistry, genetics, metabolism, clinical impli-
`cations, and commercial aspects.
`This book is intended as a resource for students and practitioners of protein
`chemistry who use albumin as a model protein for physical or chemical studies,
`as well as for clinical researchers interested in plasma protein metabolism and in
`transport of substances in the blood. I hope it will also prove useful to those
`studying genetic variation, of which much has been learned concerning albumin
`in recent years, and to molecular biologists who use albumin as a paradigm for
`elucidating the mechanisms of genetic activation and control. The largest group
`to whom this book may prove of value, however, are those who do not study
`albumin but use it for its beneficial properties. I refer to the surgeons who
`administer albumin intravenously to bolster the failing circulation of their
`patients, or the nutritionists who give albumin to promote intestinal function so
`that their patients may eat again. I refer also to the many workers in academic,
`medical, and industrial laboratories who include albumin as an essential compo-
`nent of the supporting medium of their cell cultures or who add albumin in vitro
`to protect delicate macromolecules from adsorption to the surfaces of containers.
`I hope that each of these groups will find some information pertinent to their
`
`XV
`
`MPI EXHIBIT 1042 PAGE 17
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`

`

`xvi
`
`Preface
`
`particular application and will delve a bit into the other sections of this book so
`they may gain a better overall appreciation of the properties and mysteries of the
`protein they are using. They may then be better prepared to understand the func-
`tioning of albumin in the system under study and perhaps to understand the
`functioning of the system itself.
`Various problems arise in research systems from inadequate knowledge of
`the properties of albumin. When it is added as a support protein to avoid effects
`of the surface of a container on enzymes or cells, its avidity for fatty acids and
`metals may affect the system's performance. Commercial albumin preparations
`have all been heated to 60 ~ (the degree symbol refers to degrees Celsius through-
`out this book), which can cause subtle changes in its tertiary structure; users
`should also be aware that octanoate or N-acetyltryptophan are commonly added
`to protect albumin from denaturation on heating, and traces of these ligands
`remain unless removed by special treatment. Smidgens of granulocyte proteases,
`of insulin, and of otl-antitrypsin may copurify with albumin and cause strange
`results in cell cultures. I hope that this book, without being overwhelmingly
`technical, can at least assist in a more-informed application of this protein.
`My own familiarity with albumin arose, by chance, quite early in my adven-
`tures in biochemistry while a graduate student in the laboratory of Christian B.
`Anfinsen at Harvard Medical School. This was in the immediate post-World
`War II period--radioisotopes of convenient half-life had just become available
`as a by product of the atomic pile. Only four years earlier, my college biochem-
`istry course had categorized proteins among the colloids. While measuring the
`incorporation of '4C into the proteins of a chicken liver slice system, it became
`apparent after many hours of fractionation in a subzero cold room that the most
`highly labeled protein in the system was one which had been secreted into the
`incubation medium. Naively, perhaps, I already believed that the homogeneous,
`soluble protein of about 70,000 Da was albumin, but the skeptical Dr. Anfinsen
`was only convinced by a beautiful white immune precipitate which formed
`before our eyes when an antiserum to chicken serum proteins was added.
`The proximity of the Harvard Physical Chemistry Laboratory under Edwin
`J. Cohn provided advisors such as John T. Edsall and Douglas M. Surgenor on
`the properties of albumin. It also provided the imposing E. J. Cohn himself as
`the chairman for my thesis defense, a daunting experience for an awestruck
`graduate student. This group (see Chapters 1 and 7, and particularly Fig. 1-1)
`had just completed the major wartime effort of the American Red Cross blood
`fractionation program to provide human albumin as a stable substitute for blood
`plasma for wounded soldiers on the battlefield. A later visit to the Carlsberg
`Laboratorium directed by the eminent protein chemist Kai Linderstr~m-Lang
`helped me gain an appreciatiofi of the sturdiness and resiliency of the albumin
`molecule. This appreciation grew during a period of more than three decades in
`the laboratories of The Mary Imogene Bassett Hospital, a forward-looking insti-
`
`MPI EXHIBIT 1042 PAGE 18
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`

`

`Preface
`
`xvii
`
`tution in rural upstate New York in which I was encouraged in my pursuit of this
`protein without interference. Here I was joined by colleagues Richard C. Feld-
`hoff and Roberta G. Reed, who were likewise intrigued by albumin and who
`have continued its study, Dr. Reed at the Bassett Hospital and Dr. Feldhoff at the
`University of Louisville.
`One of the joys of this pursuit has been meeting and exchanging ideas with
`leading students of albumin biochemistry. In many cases they have openly fur-
`nished unpublished information and made suggestions without personal gain.
`While the names of colleagues appear throughout this volume, I would like to
`thank in particular for their friendship and, frequently, hospitality Leon L.
`Miller, Peter N. Campbell, Julian B. Marsh, J. D. Judah, Gerhard Schreiber,
`Hans Glaumann, Colvin Redman, and Marcus A. Rothschild, researchers in
`albumin biosynthesis; Margaret J. Hunter, Walter L. Hughes, Joseph E Foster,
`Frank R. N. Gurd, Claude Lapresle, T. P. King, R. H. McMenamy, James R.
`Brown, B. Meloun, Arthur A. Spector, Rolf Brodersen, and Ingvar Sj6gren, who
`studied its chemistry; and Franco Porta, Andrew T~imoky, Stephen O. Brennan,
`Monica Galliano, Frank W. Putnam, Achilles Dugaiczyk, D. R. Schoenberg, and
`Luc B61anger, pioneers in the study of its genetic makeup.
`I hope that I have treated the reports of all of these albumin colleagues fairly
`in this book; it was certainly my intent. In addition, I express my thanks to sev-
`eral who helped in its preparation: John S. Finlayson of the U.S. Food and Drug
`Administration, who has kept an eye on commercial albumin preparations for
`years, and Jean A. Thomas and Timothy Tiemann of Miles Laboratories, Inc.,
`who have helped me understand the complexities of the commercial production
`of a protein in bulk. The willing help of the medical librarians of the Bassett
`Hospital, Linda Muehl and Robin Phillips, has been invaluable. I would be
`remiss to conclude without a word of appreciation to the many kind people who
`have stimulated and encouraged me in the world of science: Christian B. Anfin-
`sen, Eric G. Ball, and A. Baird Hastings of Harvard Medical School; Joseph W.
`Ferrebee, James Bordley, III, Clinton V. Z. Hawn, Charles A. Ashley, Roberta G.
`Reed, Gary A. Weaver, and Eugene W. Holowachuck of the Bassett Hospital;
`and also John H. Powers of that institution, who always urged me to write this
`book. For encouraging my curiosity at an earlier age, I owe a large debt to my
`mother, Miriam Lenhardt Peters, and to my first instructor of rigorous science,
`Susie Kriechbaum, high school geometry teacher. My own advice to students
`seeking a career in biological research has consistently been: Study mathematics
`and logic and the great science of chemistry. Then you will be better able to
`understand the marvelous mechanisms of life. And I hope the pursuit will be as
`enjoyable and exciting for you as it has been for me.
`
`Theodore Peters, Jr.
`
`MPI EXHIBIT 1042 PAGE 19
`
`MPI EXHIBIT 1042 PAGE 19
`
`

`

`This Page Intentionally Left Blank
`
`MPI EXHIBIT 1042 PAGE 20
`
`MPI EXHIBIT 1042 PAGE 20
`
`

`

`List of Abbreviations
`
`aa
`
`AFP
`AFM
`
`A/G
`
`ALF
`
`ANS
`
`BCG
`BCP
`BMA
`bp
`BSA
`BSP
`BW
`CD
`cDNA
`CMPF
`
`COP
`
`Da
`
`Amino acid; one-letter code
`given with Fig. 2-9
`ct-Fetoprotein
`Afamin (Lichenstein et al.,
`1994)
`Albumin/globulin ratio in
`serum
`or-Albumin (B61anger et al.,
`1994)
`1-Anilino-8-naphtho-
`sulfonic acid
`Bromcresol green
`Bromcresol purple
`Bovine mercaptalbumin
`Nucleotide base pair
`Bovine serum albuimin
`Sulfobromophthalein
`Body weight
`Circular dichroism
`Copy DNA (from mRNA)
`3-Carboxyl-4-methyl-5-
`propyl-2-furanpropanoic
`acid
`Colloid osmotic pressure,
`also oncotic pressure
`Daltons
`
`DBP
`
`DE
`DEAE
`DNP
`DTNB
`
`EGF
`ER
`ESR
`EV
`FDH
`
`FDNB
`FTIR
`GC
`GFR
`GI
`GRE
`
`GuC1
`h
`HABA
`
`HBV
`
`Vitamin D-binding protein,
`also Gc-globulin
`Distal element (genetics)
`Diethylaminoethyl
`Dinitrophenyl
`Ellman's reagent, 5,5'-
`dithiobis(2-nitrobenzoic
`acid)
`Epithelial growth factor
`Endoplasmic reticulum
`Electron spin resonance
`Extravascular
`Familial dysalbuminemic
`hyperthyroidism
`Fluorodinitrobenzene
`Fourier-transform infrared
`Gas chromatography
`Glomerular filtration rate
`Gastrointestinal
`Glucocorticoid receptor ele-
`ment (genetics)
`Guanidinium chloride
`Hour
`2-(4'-Hydroxyphenylazo)
`benzoic acid
`Hepatitis B virus
`
`xix
`
`MPI EXHIBIT 1042 PAGE 21
`
`MPI EXHIBIT 1042 PAGE 21
`
`

`

`x x
`
`List of Abbreviations
`
`HIV
`
`HMA
`HPLC
`
`HSA
`IDDM
`
`Ig
`IL
`IR
`kb
`kDa
`L
`LCFA
`
`Human immunodeficiency
`virus
`Human mercaptalbumin
`High-performance liquid
`chromatography
`Human serum albumin
`Insulin-dependent (Type-I)
`diabetes mellitus
`Immunoglobulin
`Interleukin
`Infrared
`Kilobase
`Kilodaltons
`Liter
`Long-chain fatty acids,
`C 16-C20
`Moles/liter
`M
`MCFA Medium-chain fatty acids,
`C6-C14
`Mole/mole
`M/M
`MMADS Monoacetyldiaminophenyl
`sulfone (bilrubin analog)
`Messenger ribonucleic acid
`Million years (evolution)
`Nagase analbuminemic rat
`National Ae