Improving Broad Specificity Hapten Recognition with Protein Engineering

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• 作者：Teemu Korpimä ; ki ; Jaana Rosenberg ; Pekka Virtanen ; Tuomas Karskela ; Urpo Lamminmä ; ki ; Mika Tuomola ; Markus Vehniä ; inen ; and Petri Saviranta
• 刊名：Journal of Agricultural and Food Chemistry
• 出版年：2002
• 出版时间：July 17, 2002
• 年：2002
• 卷：50
• 期：15
• 页码：4194 - 4201
• 全文大小：111K
• 年卷期：v.50,no.15(July 17, 2002)
• ISSN：1520-5118

文摘

Sulfa antibiotics (sulfonamides) are derivatives of p-aminobenzenesulfonamide that are widely usedin veterinary medicine. Foods derived from treated animals may be contaminated with these drugs.However, current immunobased sulfonamide detection methods are unfit for screening of productsbecause they are either too insensitive or specific for a few compounds only. An immunoassay capableof detecting all sulfas in a single reaction would be ideal for screening. For development of a bindercapable of binding all sulfas, a protein engineering approach was chosen and the properties ofmonoclonal antibody 27G3 were improved with mutagenesis followed by selection with phage display.Several different mutant antibodies were isolated. The cross-reaction profile of the best mutant antibodywas significantly improved over that of the wild-type antibody: it was capable of binding 9 of thetested 13 sulfonamides within a narrow concentration range and also bound the rest of the sulfas,albeit within a wider concentration range.Keywords: Drug residues; group specificity; phage display; protein engineering; sulfonamides; time-resolved fluoroimmunoassay