Design and selection of ligands for affinity chromatography

Labrou, Nikolaos E.

doi:10.1016/s1570-0232(03)00098-9

Cited by 126 publications

(87 citation statements)

References 79 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…However, the resin is very expensive (particularly when used in industrial scale chromatography columns) and can undergo degradation (41) and leach into the final product, causing toxicity (42). However, conventional chromatographic methods are suitable for large-scale processes because they are less expensive, easier to use, and largely resistant to chemical and biological degradation (43,44). Therefore, we developed a two-step strategy that exploited two intrinsic properties of IgG1 monoclonal antibodies: their pI and their ability to bind metal ions because of the presence of a surface-accessible histidine-rich region near the C terminus.…”

Section: Applied Biological Sciencesmentioning

confidence: 99%

Cost-effective production of a vaginal protein microbicide to prevent HIV transmission

Ramessar

Rademacher

Sack

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

148

124

View full text Add to dashboard Cite

A series of small-molecule microbicides has been developed for vaginal delivery to prevent heterosexual HIV transmission, but results from human clinical trials have been disappointing. Proteinbased microbicides, such as HIV-specific monoclonal antibodies, have been considered as an alternative approach. Despite their promising safety profile and efficacy, the major drawback of such molecules is the economy of large-scale production in mammalian cells, the current system of choice. Here, we show that an alternative biomanufacturing platform is now available for one of the most promising anti-HIV antibodies (2G12). Our data show that the HIV-neutralization capability of the antibody is equal to or superior to that of the same antibody produced in CHO cells. We conclude that this protein production system may provide a means to achieve microbicide ingredient manufacture at costs that would allow product introduction and manufacture in the developing world.

show abstract

Section: Applied Biological Sciencesmentioning

confidence: 99%

Cost-effective production of a vaginal protein microbicide to prevent HIV transmission

Ramessar

Rademacher

Sack

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

148

124

View full text Add to dashboard Cite

show abstract

“…When coupled with highthroughput screening, such libraries allow the identification of a potential affinity ligand for a particular protein molecule. The systematic evolution of ligands by exponential enrichment is a similar approach that creates RNA and DNA oligonucleotide libraries (aptamers) and identifies a potential affinity ligand by screening (24) . Currently, recombinant DNA technology enables the production of the desired protein with a fusion affinity tag.…”

Section: The Concept Of Affinitymentioning

confidence: 99%

Affinity-Based Strategies for Protein Purification

Mondal

Gupta²,

Roy³

2006

Anal. Chem.

View full text Add to dashboard Cite

“…As a result, the clones bound to proteins with artificial conformations when antigens were immobilized on a solid matrix were avoided (16). Although phage library-derived ligands were extensively studied (23,25,26), employing phage particles directly as affinity ligands was a relatively new concept. In the present work, phage-displayed scFv fragments were cross-linked to serve as affinity matrices for a favorable alternative, compared with phage-displayed peptides or wild-type phages as reported before (14,19,27).…”

Section: Discussionmentioning

confidence: 99%

A Novel Strategy for Proteome-wide Ligand Screening Using Cross-linked Phage Matrices

Qian

Liu

Tang

et al. 2010

Journal of Biological Chemistry

View full text Add to dashboard Cite

To find a suitable ligand from a complex antigen system is still a mission to be accomplished. Here we have explored a novel "library against proteome" panning strategy for ligand screening and antigen purification from a complex system using phage-displayed antibody technology. Human plasma proteome was targeted for phage library panning. During the process, the panning was carried out in solution, using a biotin/ streptavidin beads separation system, for three rounds. Nine monoclonal phages, bound tightly to a number of unknown plasma proteins, were selected from the last round, six of which were directly employed as cross-linked matrices to purify their corresponding antigens from the plasma. The proteins isolated by G5 and E1 matrices were identified as amyloid protein and apolipoprotein A-I precursor, respectively. The results demonstrated that it was feasible to simultaneously obtain a number of ligand phages for various antigens, including low abundant proteins in a non-comparative proteome-wide system.

show abstract

Design and selection of ligands for affinity chromatography

Cited by 126 publications

References 79 publications

Cost-effective production of a vaginal protein microbicide to prevent HIV transmission

Cost-effective production of a vaginal protein microbicide to prevent HIV transmission

Affinity-Based Strategies for Protein Purification

A Novel Strategy for Proteome-wide Ligand Screening Using Cross-linked Phage Matrices

Contact Info

Product

Resources

About