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Review: Multipoint binding and heterogeneity in immobilized metal affinity chromatography

Johnson, Robert D. and Arnold, Frances H. (1995) Review: Multipoint binding and heterogeneity in immobilized metal affinity chromatography. Biotechnology and Bioengineering, 48 (5). pp. 437-443. ISSN 0006-3592. doi:10.1002/bit.260480505.

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Studies carried out using engineered proteins clearly demonstrate that adsorption to derivatized surfaces involves multiple interactions between functional groups on the protein and complementary sites distributed on the surface. The fact that adsorption involves multipoint interactions has important implications for the design of separations processes and for the interpretation of heterogeneity in biological recognition phenomena. Increasing the density of surface metal sites (immobilized copper ions) is found to be functionally equivalent to increasing the number of metal‐coordinating groups on the protein (histidines and deporotonated amines), m in that both processes increase the likelihood of simultaneous interactions between the protein and the surface. A consequence of multiple‐site interactions is a significant in crease in protein binding affinity that depends on the arrangement of surface sites. A protein will show the highest affinity for arrangements of surface sites which best match its own pattern of functioal groups and will show lower affinity for less optimal arrangements, resulting in binding that is inherently heterogeneous. We have found that reversible protein adsorption in immobilized metal affinity chromatography (IMAC) is described by the Temikin model, which characterizes binding heterogeneity by a uniform distribution of binding energies over the population of surface binding sites.

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Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 1995 John Wiley & Sons. Manuscript received: 22 November 1994. Manuscript accepted: 19 June 1995.
Subject Keywords:chromatography; protein binding; metal affinity chromatography; heterogeneous adsorption
Issue or Number:5
Record Number:CaltechAUTHORS:20180822-140726673
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Official Citation:Johnson, R. D. and Arnold, F. H. (1995), Review: Multipoint binding and heterogeneity in immobilized metal affinity chromatography. Biotechnol. Bioeng., 48: 437-443. doi:10.1002/bit.260480505
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:89043
Deposited By: George Porter
Deposited On:22 Aug 2018 22:42
Last Modified:16 Nov 2021 00:31

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