Rapid method for measuring scFv thermal stability by yeast surface display

Brent A. Orr, Lori M. Carr, K. Dane Wittrup, Edward J Roy, David M Kranz

Research output: Contribution to journalArticle

Abstract

We have characterized a simplified method to determine the relative thermal stability of single-chain antibodies by following the irreversible denaturation of scFv fusions on the surface of yeast by flow cytometry. The method was highly reproducible and correlated well with other methods used to monitor thermal denaturation of the soluble proteins. We found a range of thermal stabilities for wild-type single-chain antibodies with half-maximum denaturation temperatures between 43 and 61°C. The ability to quantitate thermal stability of antibodies or other proteins that are immobilized on the surface of yeast allows rapid comparisons of primary structural information with stability. Thermal denaturation could be a useful parameter to consider in the choice of scFv fragments for various applications.

Original languageEnglish (US)
Pages (from-to)631-638
Number of pages8
JournalBiotechnology Progress
Volume19
Issue number2
DOIs
StatePublished - Mar 1 2003

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Hot Temperature
Yeasts
Single-Chain Antibodies
Immobilized Proteins
Protein Denaturation
Flow Cytometry
Temperature
Antibodies

ASJC Scopus subject areas

  • Biotechnology

Cite this

Rapid method for measuring scFv thermal stability by yeast surface display. / Orr, Brent A.; Carr, Lori M.; Wittrup, K. Dane; Roy, Edward J; Kranz, David M.

In: Biotechnology Progress, Vol. 19, No. 2, 01.03.2003, p. 631-638.

Research output: Contribution to journalArticle

Orr, Brent A. ; Carr, Lori M. ; Wittrup, K. Dane ; Roy, Edward J ; Kranz, David M. / Rapid method for measuring scFv thermal stability by yeast surface display. In: Biotechnology Progress. 2003 ; Vol. 19, No. 2. pp. 631-638.
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