synthetic peptides for antibody production PDF
Preview synthetic peptides for antibody production
SYNTHETIC PEPTIDES FOR ANTIBODY PRODUCTION Cover: Spatial representations of the amino acid sequences (amino acids 335-350) from lXI-antitrypsin variants. Top: the IX-helical peptide from the Z variant; Bottom: the IX-helical peptide from the M variant. The glutamic acid (M variant) and lysine (Z variant), which form the only amino acid difference in the protein, are accented in magenta. (Images made with the kind assistance of Dr. Jeanine van Rooyen and Dr. Jan Raap. RUL) SYNTHETIC PEPTIDES FOR ANTIBODY PRODUCTION SYNTHETISCHE PEPTIDEN VOOR ANTILICHAAM PRODUKTIE PROEFSCHRIFT ter verkrijging van de graad van doctor aan de Erasmus Universiteit van Rotterdam op gezag van de Rector Magnificus prof. dr. P.W.C. Akkermans M.A. en volgens het besluit van het College van Promoties. De openbare verdediging zal plaatsvinden op woensdag 29 november 1995 am 15.45 Uur door Neeltje Dina Zegers geboren te Terneuzen PROMOTIE COMMISSIE PROMOTOR: PROF.DR. E.H.J.H.M. CLAASSEN CO-PROMOTOR: DR. W.J.A. BOERSMA OVERIGE LEDEN: PROF. DR. W. VAN EWIJK PROF.DR. J.J. HAAIJMAN PROF.DR. TH.H. VAN DER KWAST CIP-DATA KONINKLIJKE BIBLIOTHEEK, DEN HAAG Zegers, NeeJtje Dina Synthetic peptides for antibody production / Neeltje Dina Zegers. - [S.I.: s.n.]. - III. Thesis Erasmus Universiteit Rotterdam. - With ref. - With summary in Dutch. ISBN 90-5412-023-1 Subject headings: peptides / antibodies / immunochemical detection. Wie alsmaar lei op de wind, komi aan zaaien /liel loe en \Vie naar de wolken blijft kijken, komi niel 101 oogslen. Begin in de morgen Ie zaaien en gun je hand 101 de avond geen rust. Je weel imll1ers niel of hel de ene keer lukl of de andere, of dal hel beide keren goed uilvall. Hel licllt is zalig en hel is een weldaad voor de ogen de zon Ie zien. Hoe lallg iemalld ook leeft, laal hij gellielen vall elke dag ell bedenken, dat er 1I0g gelloeg donkere dagen zullen Zijll en dal alles wal damna komi ijde/ is. Prediker 11: 4, 6-8 Aan mijn ouders Aan Jaap Abbreviations ABBREVIATIONS (general) AA Amino acid AFC Antibody forming cell ALL Acute lymphoblastic leukemia ApoE Apo-lipoprotein E "I-AT (Xl-antitrypsin AP Alkaline phosphatase AR Androgen honnone receptor BCIP 5-bromo-4-chloro-3-indolyl phosphate BCP Branched chain peptide BD B cell determinant BMH Bismaleimidohexane BPH Benzoic acid para hydroxylase BSA Bovine serum albumin CD Cluster of differentiation CFA Complete Freund's adjuvant CGG Chicken gamma globulin CML Chronic myeloid leukemia CS Circumsporozoite CSP Circumsporozoite protein CTL Cytolytic T cell lysis DDA Dimethyldioctadecylammonium bromide DTH Delayed type hypersensitivity EAE Experimental autoimmune encephalomyelitis EDC l-ethyl-3-(3-dimethylaminopropy l)-carbodiimide ELISA Enzyme linked immuno-sorbent assay FCS Fetal calf serum FMDV Foot-and-mouth disease vilus Fmoc 9-Fluorenylmethoxycarbonyl {3-Gal {3-galactosidase GA Glutaraldehyde Gpl20 glycoprotein 120 kDa (HIV-I) hAR Human androgen receptor HEL Hen egg lysozyme HIV-I Human immunodeficiency virus type 1 HRP Horse radish peroxidase HSA Human serum albumin HuIgG Human immunoglobulin G IEF Isoelectric focusing IFA Incomplete Freund's adjuvant 6 Abbreviations IFN-X Interferon gamma IL-# Interleukin-# i.p. Intraperitoneal ISCOM Inullune stimulating complex i. v. Intravenous KLH Keyhole limpet hemocyanin MAbs Monoclonal antibodies MAP Multiple antigen peptide MBS m-maleimidobenzoyl-N-hydroxysuccinimide ester MHC Major histocompatibility complex MLA Monophosphoryl lipid A NBT nitroblue tetrazolium NK Natural Killer OPD o-phenylendiamine PAbs Polyclonal antibodies PBS Phosphate-buffered saline PNP p-Nitrophenylphosphate PPD Purified protein derivative (of tuberculin) P3C tripalmitoyl-S-glycerylcysteine RAM-PO Rabbit anti mouse Ig conjugated with horseradish peroxidase RAM-AP Rabbit anti mouse Ig conjugated with alkaline phosphatase RI881 [17 a-methyl-'Hj methyltrienolone SAF Syntex adjuvant formulation s.c. Subcutaneous SE Staphylococcus enterotoxin SP Synthetic peptide SPDP N-succinimidyl-3-(2-pyridyldithio)-propionate t-Boc tert ia ry -bu ty I-oxycar bony I TCR T cell receptor TD T cell detemlinant Th T helper TT Tetanus toxoid TFE Trifluoroethanol TFA Trifluoroacitic acid WIO Water in oil emulsion X-Gal 5-bromo-4-chloro-3-indolyl-galactosidase 7 Abbreviations Componds used in peptide synthesis Acm Acetamidomethyl Boc Butoxycarbonyl Bop Benzotriazole-l-yl-oxy-tris(d imethylamino )-phosphoniumhexafluorophosphate 2-Br-Z 2-Bromobenzyloxycarbonyl Bzl Benzyl 2-CI-Z 2-Chlorobenzyloxycarbonyl DCC N ,N' -Dicyclohexylcarbodiimide DCM Dichloromethane DIPCDI Diiso-propylcarbodiimide Dnp 2,4-Dinitrophenyl DVB Divinylbenzene Fmoc ~-fluoreneylmethyloxycarbonyl HBTU 2-(IH-Benzotriazole-l-yl)-1, 1,3 ,3-tetramethyluronium hexatluorophosphate HMP 4-(Hydroxymethyl)phenoxymethyl HMPB 4-hydroxymethyl-3-methoxy-phenoxybutyric acid HOBt l-hydroxybenzotriazole HONSu N-hydroxysuccinimide HOOBt 3-hydrox y-3 ,4-dihydro-1 ,2, 3-benzotriazin-4-one MBHA Methylbenzylhydrylamine 4-MeBzl 4-Methylbenzyl Mts Mesitylene-2-sulphonyl OBzl Benzyloxy ODhbt 3-Hydroxy-4-oxo-3 ,4-dihydro-1 ,2,3-benzotriazine ester ONp Nitrophenyl ester ONSu N-hydroxysuccinimide ester OPfp pentafluorenyl ester OtBu t-Butoxy PAM phenylacetamidomethyl PEG Polyethyleneglycol Pmc 2,2,5,7,8-Pentamethylchroman-6-sulphonyl PSA Performed symmetrical anhydrides PyBOP Benzotriazole-l-yl-oxy-tris-pyrrolidino-phosphoniumhexatluorophosphate SPPS Solid phase peptide synthesis tBu t-Butyl TBTU 2-(lH-Benzotriazole-l-yl)-1, 1,3 ,3-tetramethyluronium tetratluoroborate TFA Trifluoracetic acid Tos Tosyl Trt Trityl t-Boc tertiair-Butoxycarbonyl Z Benzyloxycarbonyl 8 Abbreviations AMINO ACID CODES One letter Three letter Amino acid code code A Ala Alanine C Cys Cysteine D Asp Asparagic acid E Glu Glutamic acid F Phe Phenylalanine G Gly Glycine H His Histidine I lie Isoleucine K Lys Lysine L Leu Leucine M Met Methionine N Asn Asparagine P Pro Proline Q Gin Glutamine R Arg Arginine S Ser Serine T Tllr Threonine V Val Valine W Trp Tryptophan y Tyr Tyrosine 9 Con/e11ls CONTENTS Abbreviations 6 Chapter 1. General introduction 13 Chapter 2. Introduction to experimental work 51 PART 1. HOMOLOGY Chapter 3. An improved conjugation method for controlled covalent coupling of synthetic peptides to proteins using glutaraldehyde in a dialysis method 57 Chapter 4. Epitope prediction and conformation for the human androgen receptor: Generation of monoclonal antibodies for mUlti-assay performance following the synthetic peptide strategy 69 PART II. DETECTION OF VARIANT PROTEINS Chapter 5. The potency of peptides in immune responses and in immune response analysis: single amino acid specificity 91 Chapter 6. Detection of genetic variants of £xl-antitrypsin with site-specific monoclonal antibodies 117 Chapter 7. Generation and selection of monoclonal antibodies to variants of £xl-antitrypsin specific for one single amino acid using synthetic peptides: Importance of conjugation of synthetic detelminants 131 PART III. T CELL EPITOPES Chapter 8. Use of T cell epitopes in raising inmmne responses 137 Chapter 9. Peptide-induced memory (IgG) response, cross-reactive with native proteins, requires covalent linkage of a B cell epitope with a T cell epitope 169 Chapter 10 Multiple antigen peptides for immunization 181 10
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