Facile Generation of Heat-Stable Antiviral and Antitoxin Single Domain Antibodies from a Semisynthetic Llama Library
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- 作者：Ellen R. Goldman ; George P. Anderson ; Jinny L. Liu ; James B. Delehanty ; Laura J. Sherwood ; Lisa E. Osborn ; Larry B. Cummins ; Andrew Hayhurst
- 刊名：Analytical Chemistry
- 出版年：2006
- 出版时间：December 15, 2006
- 年：2006
- 卷：78
- 期：24
- 页码：8245 - 8255
- 全文大小：514K
- 年卷期：v.78,no.24(December 15, 2006)
- ISSN：1520-6882
文摘
Llamas possess a class of unconventional immunoglobulins that have only heavy chains; unpaired heavy variabledomains are responsible for antigen binding. Thesedomains have previously been cloned and expressed assingle domain antibodies (sdAbs); they comprise thesmallest known antigen binding fragments. SdAbs havebeen shown to bind antigens at >90 
C and to refold afterbeing denatured. To take advantage of the remarkableproperties of sdAbs, we constructed a large, semisyntheticllama sdAb library. This library facilitated the rapidselection of binders to an array of biothreat targets. Weselected sdAb specific for live vaccinia virus (a smallpoxvirus surrogate), hen egg lysozyme, cholera toxin, ricin,and staphylococcal enterotoxin B. The selected sdAbpossessed high specificity as well as enhanced thermalstability in comparison to conventional IgG and scFvantibodies. We also determined equilibrium dissociationconstants as well as demonstrated the use of severalantitoxin sdAbs as effective capture and reporter molecules in sandwich assays on the Luminex instrument.The ability to rapidly select such rugged antibodies willenhance the reliability of immunoassays by extendingshelf life and the capacity to function in hostile environments.
C and to refold afterbeing denatured. To take advantage of the remarkableproperties of sdAbs, we constructed a large, semisyntheticllama sdAb library. This library facilitated the rapidselection of binders to an array of biothreat targets. Weselected sdAb specific for live vaccinia virus (a smallpoxvirus surrogate), hen egg lysozyme, cholera toxin, ricin,and staphylococcal enterotoxin B. The selected sdAbpossessed high specificity as well as enhanced thermalstability in comparison to conventional IgG and scFvantibodies. We also determined equilibrium dissociationconstants as well as demonstrated the use of severalantitoxin sdAbs as effective capture and reporter molecules in sandwich assays on the Luminex instrument.The ability to rapidly select such rugged antibodies willenhance the reliability of immunoassays by extendingshelf life and the capacity to function in hostile environments.