Bioconjugating Technique英文资料.pdf

1、BioconjugateTechniques2nd EditionThis page intentionally left blankBioconjugateTechniques2nd EditionGreg T.HermansonPierce Biotechnology,Thermo Fisher Scientifi c,Rockford,Illinois,USAAMSTERDAM BOSTON HEIDELBERG LONDON NEW YORK OXFORDPARIS SAN DIEGO SAN FRANCISCO SINGAPORE SYDNEY TOKYO Academic Pres

2、s is an imprint of ElsevierThe molecular model on the cover is a nitrite reductase enzyme obtained from the RCSB Protein Data Bank(2afn),as determined by Murphy,M.E.,Turley,S.,Kukimoto,M.,Nishiyama,M.,Horinouchi,S.,Sasaki,H.,Tanokura,M.,and Adman,E.T.(1995)Structure of Alcaligenes faecalis nitrite r

3、eductase and a copper site mutant,M150E,that contains zinc.Biochemistry 34,1210712117.The space-fi lling model was created from the coordinate fi le using PovChem and the fi nal image ray-traced using POV-Ray Academic Press is an imprint of Elsevier 84 Theobalds Road,London WC1X 8RR,UK Radarweg 29,P

4、O Box 211,1000 AE Amsterdam,The Netherlands 30 Corporate Drive,Suite 400,Burlington,MA 01803,USA 525 B Street,Suite 1900,San Diego,CA 92101-4495,USA First edition 1996 Second edition 2008 Copyright 2008 Elsevier Inc.All rights reserved No part of this publication may be reproduced,stored in a retrie

5、val system or transmitted in any form or by any means electronic,mechanical,photocopying,recording or otherwise without the prior written permission of the publisher.Permissions may be sought directly from Elseviers Science&Technology Rights Department in Oxford,UK:phone(?44)(0)1865 843830;fax(?44)(

6、0)1865 853333;email:.Alternatively you can submit your request online by visiting the Elsevier web site at http:/ selecting Obtaining permission to use Elsevier material Notice No responsibility is assumed by the publisher for any injury and/or damage to persons or property as a matter of products l

7、iability,negligence or otherwise,or from any use or operation of any methods,products,instructions or ideas contained in the material herein.British Library Cataloguing in Publication Data A catalogue record for this book is available from the British Library Library of Congress Cataloging-in-Public

8、ation Data A catalog record for this book is available from the Library of Congress ISBN:978-0-12-370501-3 Typeset by Charon Tec Ltd(A Macmillan Company),Chennai,I Printed and bound in the United States of America 08 09 10 11 10 9 8 7 6 5 4 3 2 For information on all Academic Press publicationsvisit

9、 our website at For Amy and Meghan,who,since the fi rst edition was published,have now become interested in pursuing careers in microbiology and medicine.viContents Overview PART IBioconjugate Chemistry 1.Functional Targets 3 2.The Chemistry of Reactive Groups 169 PART II Bioconjugate Reagents 3.Zer

10、o-Length Crosslinkers 215 4.Homobifunctional Crosslinkers 234 5.Heterobifunctional Crosslinkers 276 6.Trifunctional Crosslinkers 336 7.Dendrimers and Dendrons 346 8.Cleavable Reagent Systems 391 9.Fluorescent Probes 396 10.Bifunctional Chelating Agents and Radioimmunoconjugates 498 11.Biotinylation

11、Reagents 506 12.Iodination Reagents 546 13.Silane Coupling Agents 562 14.Microparticles and Nanoparticles 582 15.Buckyballs,Fullerenes,and Carbon Nanotubes 627 16.Mass Tags and Isotope Tags 649 17.Chemoselective Ligation:Bioorthogonal Reagents 666 18.Discrete PEG Reagents 707 PART III Bioconjugate A

12、pplications19.Preparation of HaptenCarrier Immunogen Conjugates 745 20.Antibody Modifi cation and Conjugation 783 21.Immunotoxin Conjugation Techniques 824 22.Preparation of Liposome Conjugates and Derivatives 858 23.AvidinBiotin Systems 900 24.Preparation of Colloidal Gold-Labeled Proteins 924 25.M

13、odifi cation with Synthetic Polymers 936 26.Enzyme Modifi cation and Conjugation 961 27.Nucleic Acid and Oligonucleotide Modifi cation and Conjugation 969 28.Bioconjugation in the Study of Protein Interactions 1003viiDetailed Contents Preface to the Second Edition xxiiiPreface to the First Edition x

14、xvi Acknowledgments xxviiiHealth and Safety xxixIntellectual Property xxx PART I Bioconjugate Chemistry 1.Functional Targets 1.Modifi cation of Amino acids,Peptides,and Proteins 3 1.1.Protein Structure and Reactivity 4 Amino Acids 4 Nucleophilic Reactions and the pI of Amino Acid Side Chains 13 Seco

15、ndary,Tertiary,and Quaternary Structure 15 Prosthetic Groups,Cofactors,and Post-Translational Modifi cations 19 Protecting the Native Conformation and Activity of Proteins 21 Oxidation of Amino Acids in Proteins and Peptides 23 Solvent Accessibility of Functional Targets in Proteins 291.2.Protein Cr

16、osslinking Methods 322.Modifi cation of Sugars,Polysaccharides,and Glycoconjugates 352.1.Carbohydrate Structure and Functionality 36 Basic Sugar Structure 37 Sugar Functional Groups 39 Polysaccharide and Glycoconjugate Structure 44 2.2.Carbohydrate and Glycan Conjugation Methods 493.Modifi cation of

17、 Nucleic Acids and Oligonucleotides 503.1.Polynucleotide Structure and Functionality 51 Nucleotide Functional Groups 53 RNA and DNA Structure 623.2.Polynucleotide Crosslinking Methods 664.Creating Specifi c Functionalities 66 4.1.Introduction of Sulfhydryl Residues(Thiolation)67 Modifi cation of Ami

18、nes with 2-Iminothiolane(Traut s Reagent)67 Modifi cation of Amines with SATA 71 Modifi cation of Amines with SATP 74viiiDetailed ContentsModifi cation of Amines with SPDP 76 Modifi cation of Amines with SMPT 77 Modifi cation of Amines with N-Acetyl Homocysteine Thiolactone 80 Modifi cation of Amine

19、s with SAMSA 81 Modifi cation of Aldehydes or Ketones with AMBH 83 Modifi cation of Carboxylates or Phosphates with Cystamine 84 Modifi cation of Proteins with Cystamine 87 Modifi cation of Nucleic Acids and Oligonucleotides with Cystamine 87 Use of Disulfi de Reductants 87 Complete Reduction of Dis

20、ulfi des in Protein Molecules Using DTT 90 Use of DTT to Cleave Disulfi de-Containing Crosslinking Agents 91 Ellmans Assay for the Determination of Sulfhydryls 100 4.2.Introduction of Carboxylate Groups 101 Modifi cation of Amines with Anhydrides 102 Modifi cation of Sulfhydryls with Iodoacetate 109

21、 Modifi cation of Sulfhydryls with BMPA 111 Modifi cation of Hydroxyls with Chloroacetic Acid 113 4.3.Introduction of Primary Amine Groups 114 Modifi cation of Carboxylates with Diamines 114 Modifi cation of Sulfhydryls with N-(?-Iodoethyl)trifl uoroacetamide Aminoethyl-8 118 Modifi cation of Sulfhy

22、dryls with Ethylenimine 119 Modifi cation of Sulfhydryls with 2-Bromoethylamine 120 Modifi cation of Sulfhydryls with 2-Aminoethyl-2?-aminoethanethiolsulfonate 121 Modifi cation of Carbohydrates with Diamines 122 Modifi cation of Alkylphosphates with Diamines 124 Modifi cation of Aldehydes with Ammo

23、nia or Diamines 124 Introduction of Arylamines on Phenolic Compounds 125 Amine Detection Reagents 127 4.4.Introduction of Aldehyde Residues 129 Periodate Oxidation of Glycols and Carbohydrates 130 Oxidase Modifi cation of Sugar Residues 131 Modifi cation of Amines with NHS-Aldehydes(SFB and SFPA)132

24、 Modifi cation of Amines with Glutaraldehyde 134 Periodate Oxidation of N-Terminal Serine or Threonine Residues 136 4.5.Introduction of Hydrazine or Hydrazide Functionalities 139 Modifi cation of Aldehydes with Bis-Hydrazide Compounds 140 Modifi cation of Carboxylates with Bis-Hydrazide Compounds 14

25、2 Modifi cation of Amines with SANH,SHNH,or SHTH 143 Modifi cation of Alkylphosphates with Bis-Hydrazide Compounds 146 4.6.Introduction of Saccharide or Glycan Groups 147 Modifi cation of Amines with Mono(lactosylamido)mono(succinimidyl)suberate 149 Modifi cation of Amine or Hydrazide Molecules by C

26、arbohydrates and Glycans 150 Labeling Glycans with Fluorescent 2-Aminopyridine,2-Amino Benzamide,or Anthranilic Acid 153 Synthesis of Glycosylamines for Conjugating Glycans 155 5.Blocking Specifi c Functional Groups 156 5.1.Blocking Amine Groups 157 Detailed Contents ixSulfo-NHS Acetate 157Acetic An

27、hydride 158Citraconic Anhydride 159Maleic Anhydride 1595.2.Blocking Sulfhydryl Groups 160N-Ethyl Maleimide 160Iodoacetate Derivatives 161Sodium Tetrathionate 161Methyl Methanethiosulfonate 163 Ellmans Reagent 164Dipyridyl Disulfi de Reagents 1655.3.Blocking Aldehyde Groups 166Reductive Amination wit

28、h Tris or Ethanolamine 1665.4.Blocking Carboxylate Groups 167Tris or Ethanolamine plus EDC 167 2.The Chemistry of Reactive Groups 1.Amine Reactions 1691.1.Isothiocyanates 1701.2.Isocyanates 1701.3.Acyl Azides 1711.4.NHS Esters 1711.5.Sulfonyl Chlorides 1731.6.Aldehydes and Glyoxals 1731.7.Epoxides a

29、nd Oxiranes 1741.8.Carbonates 1751.9.Arylating Agents 1751.10.Imidoesters 1761.11.Carbodiimides 1761.12.Anhydrides 1781.13.Fluorophenyl Esters 1791.14.Hydroxymethyl Phosphine Derivatives 1801.15.Guanidination of Amines 1812.Thiol Reactions 1822.1.Haloacetyl and Alkyl Halide Derivatives 1822.2.Maleim

30、ides 1832.3.Aziridines 1842.4.Acryloyl Derivatives 1842.5.Arylating Agents 1852.6.Thiol-Disulfi de Exchange Reagents 185Pyridyl Disulfi des 186 TNB-Thiol 187xDetailed ContentsDisulfi de Reductants 187 2.7.Vinylsulfone Derivatives 188 2.8.Metal-Thiol Dative Bonds 188 2.9.Native Chemical Ligation 191

31、2.10.Cisplatin Modifi cation of Methionine and Cysteine 192 3.Carboxylate Reactions 192 3.1.Diazoalkanes and Diazoacetyl Compounds 193 3.2.Carbonyldiimidazole 194 3.3.Carbodiimides 195 4.Hydroxyl Reactions 195 4.1.Epoxides and Oxiranes 195 4.2.Carbonyldiimidazole 196 4.3.N,N?-Disuccinimidyl Carbonat

32、e or N-Hydroxysuccinimidyl Chloroformate 196 4.4.Oxidation with Periodate 197 4.5.Enzymatic Oxidation 198 4.6.Alkyl Halogens 198 4.7.Isocyanates 199 5.Aldehyde and Ketone Reactions 200 5.1.Hydrazine Derivatives 200 5.2.Schiff Base Formation 200 5.3.Reductive Amination 201 5.4.Mannich Condensation 20

33、1 6.Active Hydrogen Reactions 202 6.1.Diazonium Derivatives 202 6.2.Mannich Condensation 203 6.3.Iodination Reactions 203 7.Photo-Chemical Reactions 204 7.1.Aryl Azides and Halogenated Aryl Azides 204 7.2.Benzophenones 205 7.3.Anthraquinones 205 7.4.Certain Diazo Compounds 207 7.5.Diazirine Derivatives 208 7.6.Psoralen Compounds 208 8.Cycloaddition Reactions 210 8.1.DielsAlder Reaction 210 8.2.Com