Artificial antibodies for affinity chromatography of homologous proteins: Application to blood clotting proteins

Huiping Wu, Gaddam N. Goud, Michael Sierks

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

A method to readily isolate antibodies that bind to only one member of a family of homologous proteins is described. A library of different single chain antibody fragments can be displayed on the surface of a bacteriophage vector. Individual antibodies from this library recognizing a particular protein from a family of homologous proteins can be readily isolated by a two-step affinity screening process. In the first step antibodies which bind specifically to the undesired proteins or to homologous regions of the proteins are removed. In the second step, those antibodies specifically recognizing the desired protein are then isolated. Using this procedure and starting with a naive antibody library, a single chain antibody fragment specific to the blood clotting protein, Protein C, which did not recognize either of the homologous proteins, Factor IX or Factor X, was isolated. Similarly an antibody specific to Factor IX, but not Factor X or Protein C, was also isolated. The isolated antibodies can be readily produced, purified, and affixed to sepharose beads for affinity chromatography of the blood clotting factors. One of the key advantages to this procedure over conventional monoclonal antibody isolation is that the antibodies are isolated and produced in vitro so a broad range of related proteins, toxins, viruses, or other products can be targeted.

Original languageEnglish (US)
Pages (from-to)496-499
Number of pages4
JournalBiotechnology Progress
Volume14
Issue number3
DOIs
StatePublished - May 1998
Externally publishedYes

Fingerprint

blood coagulation
Antibody Affinity
Blood Coagulation
affinity chromatography
Affinity Chromatography
Blood Proteins
Antibodies
antibodies
Proteins
proteins
Single-Chain Antibodies
Factor X
Immunoglobulin Fragments
Factor IX
Protein C
Blood Coagulation Factors
antibody affinity
Bacteriophages
Sepharose
Libraries

ASJC Scopus subject areas

  • Food Science
  • Biotechnology
  • Microbiology

Cite this

Artificial antibodies for affinity chromatography of homologous proteins : Application to blood clotting proteins. / Wu, Huiping; Goud, Gaddam N.; Sierks, Michael.

In: Biotechnology Progress, Vol. 14, No. 3, 05.1998, p. 496-499.

Research output: Contribution to journalArticle

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