AN OVERVIEW OF AFFINITY CHROMATOGRAPHY
1 AN OVERVIEW OF AFFINITY CHROMATOGRAPHY 2 WEAK AFFINITY CHROMATOGRAPHY 3 FLUIDIZED-BED RECEPTOR–AFFINITY CHROMATOGRAPHY 4 SITE-SPECIFIC IMMOBILIZATION OF ANTIBODIES TO PROTEIN G-DERIVATIZED SOLID SUPPORTS 5 AFFINITY PURIFICATION OF MONOCLONAL ANTIBODIES 6 PROTEIN A MIMETIC (PAM) AFFINITY CHROMATOGRAPHY IMMUNOGLOBULINS PURIFICATION 7 PERIODATE OXIDATION OF ANTIBODIES FOR SITE-SELECTIVE IMMOBILIZATION IN IMMUNOAFFINITY CHROMATOGRAPHY 8 MINI-ANTIBODY AFFINITY CHROMATOGRAPHY OF LYSOZYME 9 NITROCELLULOSE-BASED IMMUNOAFFINITY CHROMATOGRAPHY 10 IMMUNOAFFINITY CHROMATOGRAPHY CONTENTS 11 AFFINITY PARTITIONING OF PROTEINS 12 BORONATE AFFINITY CHROMATOGRAPHY 13 DYE–LIGAND AFFINITY CHROMATOGRAPHY FOR PROTEIN SEPARATION AND PURIFICATION 14 DNA AFFINITY CHROMATOGRAPHY 15 AFFINITY CHROMATOGRAPHY OF PYROGENS 16 WESTERN CROSS BLOT 17 AFFINITY PERFUSION CHROMATOGRAPHY 18 PHAGE DISPLAY TECHNOLOGY AFFINITY SELECTION BY BIOPANNING 19 PHAGE DISPLAY TECHNOLOGY IDENTIFICATION OF PEPTIDES AS MODEL LIGANDS FOR AFFINITY CHROMATOGRAPHY AN OVERVIEW OF AFFINITY CHROMATOGRAPHY 1 INTRODUCTION AFFINITY CHROMATOGRAPHY IS PERVASIVELY ACCEPTED AND USED AS A TOOL IN BIO- MEDICAL RESEARCH AND BIOTECHNOLOGY; YET ITS ORIGINS ONLY 30 YEARS AGO SOME- TIMES SEEM DIMMED IN HISTORY HOWEVER, THE POTENTIAL OF THIS TECHNOLOGY CONTINUES TO STIMULATE CONTINUED DEVELOPMENT AND NEW APPLICATIONS HAVING A NEW BOOK ON THIS METHODOLOGY IS EMINENTLY APPROPRIATE TODAY AND BEING ABLE TO INTRODUCE THIS BOOK IS OUR PLEASURE AFFINITY CHROMATOGRAPHY AS IT IS KNOWN TODAY WAS INTRODUCED IN 1968 BY PEDRO CUATRECASAS, CHRIS ANFINSEN, AND MEIR WILCHEK, ONE OF THE AUTHORS OF THIS CHAPTER THOUGH FEW RELATED METHODS WERE DESCRIBED EARLIER, THE CONCEPT AND IMMENSE POWER OF BIORECOGNITION AS A MEANS OF PURIFICATION WAS INTRO- DUCED FIRST IN THAT 1968 PAPER (1) ENTITLED “AFFINITY CHROMATOGRAPHY ” IF YOU EXAMINE THE MEDLINE DATABASE FOR HOW MANY TIMES “AFFINITY CHROMA- TOGRAPHY” HAS APPEARED IN THE TITLE OF SCIENTIFIC PAPERS, YOU WILL FIND ALMOST 30,000 PAPERS CITED THIS MEANS THAT, OVER THE PAST 30 YEARS, THREE PUBLISHED PAPERS PER DAY HAVE FEATURED THIS TECHNOLOGY MOREOVER, 300 PATENTS HAVE BEEN GRANTED DURING THE LAST 2 YEARS ALONE IN A RECENT REVIEW (2), CHRIS LOWE STATED THAT AFFINITY CHROMATOGRAPHY IS A TECHNIQUE USED IN 60% OF ALL PURIFICATION PROTOCOLS SO WHAT EXACTLY IS AFFINITY CHROMATOGRAPHY—THE TECHNIQUE TO WHICH THIS BOOK IS DEVOTED? 2 AFFINITY CHROMATOGRAPHY AND ITS APPLICATIONS FOR PURIFICATION AFFINITY CHROMATOGRAPHY IS BASED ON MOLECULAR RECOGNITION IT IS A RELATIVELY SIMPLE PROCEDURE ANY GIVEN BIOMOLECULE THAT ONE WISHES TO PURIFY USUALLY HAS AN INHERENT RECOGNITION SITE THROUGH WHICH IT CAN RECOGNIZE A NATURAL OR ARTIFI- CIAL MOLECULE IF ONE OF THESE RECOGNITION PARTNERS IS IMMOBILIZED ON A POLY- AN OVERVIEW OF AFFINITY CHROMATOGRAPHY MERIC CARRIER, IT CAN BE USED TO CAPTURE SELECTIVELY THE BIOMOLECULE BY SIMPLY PASSING AN APPROPRIATE CELL EXTRACT CONTAINING THE LATTER THROUGH THE COLUMN THE DESIRED BIOMOLECULE CAN THEN BE ELUTED BY CHANGING EXTERNAL CONDITIONS, E G , PH, IONIC STRENGTH, SOLVENTS, AND TEMPERATURE, SO THAT THE COMPLEX BETWEEN THE BIOMOLECULE AND ITS PARTNER WILL NO LONGER BE STABLE, AND THE DESIRED MOL- ECULE WILL BE ELUTED IN A PURIFIED FORM NUMEROUS BOOKS AND REVIEWS ON THE APPLICATION AND THEORY OF AFFINITY CHRO- MATOGRAPHY HAVE APPEARED IN RECENT YEARS (3) HERE, WE SIMPLY LIST CLASSES OF COMPOUNDS PURIFIED BY THIS METHOD (SEE TABLE 1) 3 TECHNIQUES THAT STEM FROM AFFINITY CHROMATOGRAPHY THE BROAD SCOPE OF THE VARIOUS APPLICATIONS OF AFFINITY CHROMATOGRAPHY HAS GENERATED THE DEVELOPMENT OF SUBSPECIALTY ADAPTATIONS, MANY OF WHICH ARE NOW RECOGNIZED BY THEIR OWN NOMENCLATURE AS AN EXPRESSION OF THEIR GENERALITY AND UNIQUENESS BECAUSE SOME OF THESE APPLICATIONS HAVE A CHAPTER OF THEIR OWN IN THIS VOLUME, WE ONLY SUMMARIZE THEM IN TABLE 2 AS THIS BOOK SHOWS, SOME OF THE SUBCATEGORIES HAVE BECOME GENERALLY ACCEPTED AS USEFUL TECHNIQUES AMONG THE MOST POPULAR OF THESE AFFINITY-DERIVED TECHNIQUES IS IMMUNOAFFINITY CHROMATOGRAPHY, WHICH UTILIZES ANTIBODY COLUMNS TO PURIFY ANTIGENS, OR ANTIGEN COLUMNS TO PURIFY ANTIBODIES IMMUNOAFFINITY CHROMATOGRA- PHY IS, IN FACT, USED IN MOST BIOLOGICAL STUDIES OTHER METHODS, SUCH AS METAL– CHELATE AFFINITY CHROMATOGRAPHY, APPLY SITE-DIRECTED MUTAGENESIS TO INTRODUCE VARIOUS AFFINITY TAGS OR TAILS TO THE BIOMOLECULE TO BE PURIFIED FOR EXAMPLE, THE HIS-TAG IS USED BOTH IN METAL–CHELATE CHROMATOGRAPHY AND AS AN ANTIGEN IN IMMUNOAFFINITY CHROMATOGRAPHY MORE RECENTLY, THE USE OF COMBINATORIAL LIBRARIES HAS BECOME INCREASINGLY POPULAR FOR DEVELOPING NEW AFFINITY LIGANDS 4 CARRIERS IT IS INTERESTING THAT IN ALL THESE DEVELOPMENTS THE CARRIERS USED WERE POLYSAC- CHARIDES, MODIFIED POLYSACCHARIDES, SILICA AND TO A LESSER EXTENT POLYSTYRENE TABLE BIOMOLECULES PURIFIED BY AFFINITY CHROMATOGRAPHY 1 ANTIBODIES AND ANTIGENS 9 LECTINS AND GLYCOPROTEINS 2 ENZYMES AND INHIBITORS 10 RNA AND DNA (GENES) 3 REGULATORY ENZYMES 11 BACTERIA 4 DEHYDROGENASES 12 VIRUSES AND PHAGES 5 TRANSAMINASES 13 CELLS 6 HORMONE-BINDING PROTEINS 14 GENETICALLY ENGINEERED PROTEINS 7 VITAMIN-BINDING PROTEINS 15 OTHERS 8 RECEPTORS AN OVERVIEW OF AFFINITY CHROMATOGRAPHY EVEN TODAY, 95% OF ALL AFFINITY PURIFICATION METHODS INVOLVE AGAROSE- SEPHAROSE, THE CARRIER THAT WAS ORIGINALLY INTRODUCED IN THE FIRST PAPER ON AFFIN- ITY CHROMATOGRAPHY 5 ACTIVATION AND COUPLING IN THIS BOOK, MOST OF THE CHAPTERS DEAL WITH APPLICATION AND NOT WITH METH- ODOLOGY FOR THE PREPARATION OF THE AFFINITY COLUMNS INDEED, THE METHODOLOGY IS WELL DOCUMENTED AND WIDELY USED (4) HERE WE DESCRIBE ONLY BRIEFLY SOME OF THE PROCEDURES USED TO PREPARE AN AFFINITY COLUMN AFFINITY CHROMATOGRAPHY IS A FIVE-STEP PROCESS, WHICH CONSISTS OF ACTIVATION OF THE MATRIX, FOLLOWED BY COUPLING OF LIGANDS, ADSORPTION OF THE PROTEIN, ELU- TION, AND REGENERATION OF THE AFFINITY MATRIX A SHORT DESCRIPTION OF THE ACTIVA- TION AND COUPLING IS DESCRIBED AS FOLLOWS IN MOST STUDIES, THE ACTIVATION PROCESS IS STILL PERFORMED USING THE CYANOGEN BROMIDE METHOD HOWEVER, STUDIES ON THE MECHANISM OF ACTIVATION WITH CNBR REVEALED THAT THE USE OF THIS METHOD CAN CAUSE SERIOUS PROBLEMS THEREFORE, NEW ACTIVATION METHODS WERE DEVELOPED THAT GAVE MORE STABLE PRODUCTS THE NEWER METHODS HAVE MAINLY BEEN BASED ON CHLOROFORMATES, CARBONATES, SUCH AS N-HYDROXYSUCCINIMIDE CHLOROFORMATE OR CARBONYL BIS-IMIDAZOLE OR CARBONYL (BIS-N-HYDROXYSUCCINIMIDE) AND HYDROXYSUCCINIMIDE ESTERS, WHICH AFTER REAC- TION WITH AMINES RESULT IN STABLE CARBAMATES OR AMIDES (5,6) THE COUPLING OF LIGANDS OR PROTEINS TO THE ACTIVATED CARRIER IS USUALLY PERFORMED AT A PH SLIGHTLY ABOVE NEUTRAL DETAILS REGARDING SUBSEQUENT STEPS CAN BE FOUND IN MANY OF THE OTHER CHAPTERS OF THIS VOLUME TABLE VARIOUS TECHNIQUES DERIVED FROM AFFINITY CHROMATOGRAPHY 1 IMMUNOAFFINITY CHROMATOGRAPHY 13 AFFINITY DENSITY PERTURBATION 2 HYDROPHOBIC CHROMATOGRAPHY 14 PERFUSION AFFINITY CHROMATOGRAPHY 3 HIGH PERFORMANCE AFFINITY 15 CENTRIFUGED AFFINITY CHROMATOGRAPHY CHROMATOGRAPHY 16 AFFINITY REPULSION CHROMATOGRAPHY 4 LECTIN AFFINITY CHROMATOGRAPHY 17 AFFINITY TAILS CHROMATOGRAPHY 5 METAL-CHELATE AFFINITY CHROMATOGRAPHY 18 THEOPHILIC CHROMATOGRAPHY 6 COVALENT AFFINITY CHROMATOGRAPHY 19 MEMBRANE-BASED AFFINITY 7 AFFINITY ELECTROPHORESIS CHROMATOGRAPHY 8 AFFINITY CAPILLARY ELECTROPHORESIS 20 WEAK AFFINITY CHROMATOGRAPHY 9 DYE-LIGAND AFFINITY CHROMATOGRAPHY 21 RECEPTOR AFFINITY CHROMATOGRAPHY 10 AFFINITY PARTITIONING 22 AVIDIN-BIOTIN IMMOBILIZED SYSTEM 11 FILTER AFFINITY TRANSFER 23 MOLECULAR IMPRINTING AFFINITY CHROMATOGRAPHY 24 LIBRARY-DERIVED AFFINITY LIGANDS 12 AFFINITY PRECIPITATION AN OVERVIEW OF AFFINITY CHROMATOGRAPHY 6 RECOGNITION FIDELITY AND ANALYTICAL AFFINITY CHROMATOGRAPHY AFFINITY CHROMATOGRAPHY IS BASED ON THE ABILITY OF AN AFFINITY COLUMN TO MIMIC THE RECOGNITION OF A SOLUBLE LIGAND SUCH FIDELITY ALSO HAS PRESENTED A VEHICLE TO ANALYZE ISOCRATIC ELUTION OF A BIOLOGICAL MACROMOLECULE ON AN IMMOBILIZED LIGAND AFFINITY SUPPORT UNDER NONCHAOTROPIC BUFFER CONDITIONS ALLOWS A DYNAMIC EQUILIB- RIUM BETWEEN ASSOCIATION AND DISSOCIATION IT IS DIRECTLY DEPENDENT ON THE EQUILIB- RIUM CONSTANT FOR THE IMMOBILIZED LIGAND—MACROMOLECULE INTERACTION HENCE, AFFINITY IS REFLECTED IN THE ELUTION VOLUME THE ANALYTICAL USE OF AFFINITY CHROMATOG- RAPHY WAS DEMONSTRATED WITH STAPHYLOCOCCAL NUCLEASE (7), ON THE SAME KIND OF AFFINITY SUPPORT AS USED PREPARATIVELY (1) BUT UNDER CONDITIONS THAT ALLOWED ISOCRATIC ELUTION SIMILAR FINDINGS HAVE BEEN REPORTED BY NOW IN MANY OTHER SYSTEMS (8) OF PARTICULAR NOTE, INTERACTION ANALYSIS ON AFFINITY COLUMNS CAN BE ACCOMPLISHED OVER A WIDE RANGE OF AFFINITY, AS WELL AS SIZE OF BOTH IMMOBILIZED AND MOBILE INTERACTORS THIS ANALYSIS CAN BE ACHIEVED ON A MICROSCALE DEPENDENT ONLY ON THE LIMITS OF DETECTABILITY OF THE INTERACTOR ELUTING FROM THE AFFINITY COLUMN 7 AUTOMATION AND RECOGNITION BIOSENSORS THE ANALYTICAL USE OF IMMOBILIZED LIGANDS HAS BEEN ADAPTED TO METHODOLOGI- CAL CONFIGURATIONS WHICH ALLOW FOR AUTOMATION AND EXPANDED INFORMATION AN EARLY INNOVATION OF ANALYTICAL AFFINITY CHROMATOGRAPHY WAS ITS ADAPTATION TO HIGH- PERFORMANCE LIQUID CHROMATOGRAPHY HIGH PERFORMANCE ANALYTICAL AFFINITY CHRO- MATOGRAPHY (9) PROVIDES A RAPID MACROMOLECULAR RECOGNITION ANALYSIS AT MICROSCALE LEVEL, USING MULTIPLE POSTCOLUMN MONITORING DEVICES TO INCREASE THE INFORMATION LEARNED ABOUT ELUTING MOLECULES SIMULTANEOUS MULTIMOLECULAR ANALYSIS IS ALSO FEASIBLE, E G , BY WEAK ANALYTICAL AFFINITY CHROMATOGRAPHY (10) YEARS SINCE THE DEVELOPMENT OF AFFINITY CHROMATOGRAPHIC RECOGNITION ANALY- SIS WITH IMMOBILIZED LIGANDS FOLLOWED THE EVOLUTION OF MOLECULAR BIOSENSORS ULTIMATELY, A TECHNOLOGICAL BREAKTHROUGH FOR DIRECT INTERACTION ANALYSIS WAS THE SURFACE PLASMON RESONANCE (SPR) BIOSENSOR DEVELOPED BY PHARMACIA, CALLED BIACORE™, IN WHICH THE IMMOBILIZED LIGAND IS ATTACHED TO A DEXTRAN LAYER ON A GOLD SENSOR CHIP THE INTERACTION OF MACROMOLECULES PASSING OVER THE CHIP THROUGH A FLOW CELL IS DETECTED BY CHANGES OF REFRACTIVE INDEX AT THE GOLD SURFACE USING SPR (11,12) THE SPR BIOSENSOR IS SIMILAR IN CONCEPT TO ANALYTICAL AFFINITY CHROMATOGRA- PHY: BOTH INVOLVE INTERACTION ANALYSIS OF MOBILE MACROMOLECULES FLOWING OVER SURFACE-IMMOBILIZED LIGANDS THE SPR BIOSENSOR ALSO PROVIDES SOME UNIQUE ADVANTAGES THESE INCLUDE (1) ACCESS TO ON- AND OFF-RATE ANALYSIS, THUS PROVID- ING DEEPER CHARACTERIZATION OF MOLECULAR MECHANISMS OF BIOMOLECULAR RECOG- NITION AND TOOLS TO GUIDE THE DESIGN OF NEW RECOGNITION MOLECULES; AND (2) ANALYSIS IN REAL TIME, THUS PROMISING THE POTENTIAL TO STIMULATE AN OVERALL ACCEL- ERATION OF MOLECULAR DISCOVERY AN OVERVIEW OF AFFINITY CHROMATOGRAPHY IN ADDITION TO BIACORE, AN EVANESCENT WAVE BIOSENSOR FOR MOLECULAR RECOGNI- TION ANALYSIS HAS BEEN INTRODUCED RECENTLY BY FISONS, CALLED IASYS™ (13,14) INSTEAD OF PASSING THE ANALYTE OVER THE SENSOR CHIP THROUGH A FLOW CELL, IASYS USES A REINSERTABLE MICROCUVET SAMPLE CELL, WHICH CONTAINS INTEGRATED OPTICS A STIRRER IN THE CUVETTE ENSURES EFFICIENT MIXING TO LIMIT MASS TRANSPORT DEPENDENCE AUTOMATION IN THE ANALYTICAL USE OF IMMOBILIZED LIGANDS SEEMS LIKELY TO CONTINUE TO EVOLVE ANALYTICAL AFFINITY CHROMATOGRAPHY INCREASINGLY IS BEING ADAPTED TO SOPHISTICATED INSTRUMENTATION AND HIGH-THROUGHPUT AFFINITY SUPPORTS IN ADDITION, NEW METHODOLOGICAL CONFIGURATIONS WITH BIOSENSORS ARE BEING DEVELOPED THESE ADVANCES PROMISE TO EXPAND GREATLY THE ACCESSIBILITY OF BOTH EQUILIBRIUM AND KINETIC DATA FOR BASIC AND BIOTECHNOLOGICAL RESEARCH 8 CONCLUSIONS LOOKING BACK, AFFINITY CHROMATOGRAPHY HAS MADE A SIGNIFICANT CONTRIBUTION TO THE RAPID PROGRESS WHICH WE HAVE WITNESSED IN BIOLOGICAL SCIENCE OVER THE LAST YEARS AFFINITY CHROMATOGRAPHY, DUE TO ITS INTERDISCIPLINARY NATURE, HAS ALSO INTRO- DUCED ORGANIC, POLYMER AND BIOCHEMISTS TO THE EXCITING FIELD OF SOLVING PROBLEMS WHICH ARE PURELY BIOLOGICAL IN NATURE THUS, AFFINITY CHROMATOGRAPHY, AND THE AFFINITY TECHNOLOGIES IT HAS INSPIRED, CONTINUE TO MAKE A POWERFUL IMPACT IN FOSTER- ING THE DISCOVERY OF BIOLOGICAL MACROMOLECULES AND THE ELUCIDATION OF MOLECULAR MECHANISMS OF INTERACTION UNDERLYING THEIR BIOACTIVITIES REFERENCES 1 CUATRECASAS, P , WILCHEK, M , AND ANFINSEN, C B (1968) SELECTIVE ENZYME PURI- FICATION BY AFFINITY CHROMATOGRAPHY PROC NATL ACAD SCI USA 61, 636–643 2 LOWE, C R (1996) ADV MOL CELL BIOL 15B, 513–522 3 KLINE, T , ED (1993) HANDBOOK OF AFFINITY CHROMATOGRAPHY, MARCEL DEKKER, NEW YORK 4 WILCHEK, M , MIRON, T , AND KOHN, J (1984) AFFINITY CHROMATOGRAPHY METHODS ENZYMOL 104, 3–56 5 WILCHEK, M AND MIRON, T (1985) APPL BIOCHEM BIOTECH 11, 191–193 6 WILCHEK, M , KNUDSEN, K L AND MIRON, T (1994) IMPROVED METHOD FOR PREPAR- ING N-HYDROXYSUCCINIMIDE ESTER-CONTAINING POLYMERS FOR AFFINITY CHROMATOGRA- PHY BIOCONJUG CHEM 5, 491–492 7 DUNN, B M AND CHAIKEN, I M (1974) QUANTITATIVE AFFINITY CHROMATOGRAPHY DETERMINATION OF BINDING CONSTANTS BY ELUTION WITH COMPETITIVE INHIBITORS PROC NATL ACAD SCI 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, STEWART, W J , MAULE, C H , MOLLOY, J , AND GODDARD, N J (1993) THE RESONANT MIRROR: A NOVEL OPTICAL BIOSENSOR FOR DIRECT SENSING OF BIOMOLECULAR INTERACTIONS PART I PRINCIPLE OF OPERATION AND ASSOCIATED INSTRU- MENTATION BIOSENSORS BIOELECTRONICS 8, 347–353 14 BUCKLE, P E , DAVIES, R J , KINNING, T , YEUNG, D , EDWARDS, P R , POLLARD- KNIGHT, D , AND LOWE, C R (1993) THE RESONANT MIRROR: A NOVEL OPTICAL BIOSEN- SOR FOR DIRECT SENSING OF BIOMOLECULAR INTERACTIONS PART II APPLICATIONS BIOSENSORS BIOELECTRONICS 8, 355–363 AN OVERVIEW OF AFFINITY CHROMATOGRAPHY