Engineered protein A for the orientational control of immobilized proteins

Colin P. Johnson, Irene E. Jensen, Anil Prakasam, Ravi Vijayendran, Deborah E Leckband

Research output: Contribution to journalArticle

Abstract

This work describes the genetic engineering and characterization of a histidine-tagged fragment of protein A. The histidine tag results in the site-selective immobilization of the protein A receptor and the preservation of its high ligand affinity when immobilized on solid supports. The fragment was expressed at high yield in E. coli and purified to homogeneity. When selectively immobilized to histidine binding matrices, the protein A fragment exhibits high affinity for soluble IgG. We further demonstrate from adsorption isotherms that the receptor exhibits a homogeneous, high affinity population at densities where steric crowding between large ligands does not affect the apparent receptor affinity. This engineered receptor is appropriate for a range of applications including sensor design or those using immobilized Fc-tagged proteins.

Original languageEnglish (US)
Pages (from-to)974-978
Number of pages5
JournalBioconjugate Chemistry
Volume14
Issue number5
DOIs
StatePublished - Sep 1 2003

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Immobilized Proteins
Staphylococcal Protein A
Histidine
Proteins
Ligands
Genetic engineering
Genetic Engineering
Population Density
Adsorption isotherms
Immobilization
Escherichia coli
Adsorption
Immunoglobulin G
Sensors

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

Cite this

Engineered protein A for the orientational control of immobilized proteins. / Johnson, Colin P.; Jensen, Irene E.; Prakasam, Anil; Vijayendran, Ravi; Leckband, Deborah E.

In: Bioconjugate Chemistry, Vol. 14, No. 5, 01.09.2003, p. 974-978.

Research output: Contribution to journalArticle

Johnson, Colin P. ; Jensen, Irene E. ; Prakasam, Anil ; Vijayendran, Ravi ; Leckband, Deborah E. / Engineered protein A for the orientational control of immobilized proteins. In: Bioconjugate Chemistry. 2003 ; Vol. 14, No. 5. pp. 974-978.
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