Development of sulfhydryl-reactive silica for protein immobilization in high-performance affinity chromatography.

Development of sulfhydryl-reactive silica for protein immobilization in high-performance affinity chromatography. Research Abstract Details 

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Development of sulfhydryl-reactive silica for protein immobilization in high-performance affinity chromatography. Abstract Text:

Rangan Mallik,Chunling Wa,David S Hage,

Two techniques were developed for the immobilization of proteins and other ligands to silica through sulfhydryl groups. These methods made use of maleimide-activated silica (the SMCC method) or iodoacetyl-activated silica (the SIA method). The resulting supports were tested for use in high-performance affinity chromatography by employing human serum albumin (HSA) as a model protein. Studies with normal and iodoacetamide-modified HSA indicated that these methods had a high selectivity for sulfhydryl groups on this protein, which accounted for the coupling of 77-81% of this protein to maleimide- or iodoacetyl-activated silica. These supports were also evaluated in terms of their total protein content, binding capacity, specific activity, nonspecific binding, stability, and chiral selectivity for several test solutes. HSA columns prepared using maleimide-activated silica gave the best overall results for these properties when compared to HSA that had been immobilized to silica through the Schiff base method (i.e., an amine-based coupling technique). A key advantage of the supports developed in this work is that they offer the potential of giving greater site-selective immobilization and ligand activity than amine-based coupling methods. These features make these supports attractive in the development of protein columns for such applications as the study of biological interactions and chiral separations.

Development of sulfhydryl-reactive silica for protein immobilization in high-performance affinity chromatography. Publishing Authors By Initials

R Mallik,C Wa,DS Hage,

For similar natural sciences: chemistry: chemistry, physical: surface properties research abstracts see: natural sciences: chemistry: chemistry, physical: surface properties research

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Development of sulfhydryl-reactive silica for protein immobilization in high-performance affinity chromatography. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Analytical chemistry

VOLUME: 79

Page Numbers: 1411-24

Journal Abbreviation: Anal. Chem.

ISSN: 0003-2700

DAY: 15

MONTH: Feb

YEAR: 2007

Development of sulfhydryl-reactive silica for protein immobilization in high-performance affinity chromatography. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 370536

Development of sulfhydryl-reactive silica for protein immobilization in high-performance affinity chromatography. Keywords Mesh Terms:

KEYWORDS: Surface Properties

MESH TERMS: chemistry

Chemical & Substance for Abstract: Development of sulfhydryl-reactive silica for protein immobilization in high-performance affinity chromatography. Information

Substance Name: Silicon Dioxide

Registry Number: 7631-86-9

Grant and Affiliation Information for Development of sulfhydryl-reactive silica for protein immobilization in high-performance affinity chromatography.

AFFILIATION: Chemistry Department, University of Nebraska, Lincoln, Nebraska 68588-0304, USA.

Country: United States

AGENCY: United States NCRR

GRANT: RR015468-001

ACRONYM: RR

MEDLINETA: Anal Chem

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