Selection and Characterization of Phage Miniantibodies to Actins of Different Origin

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N. V. Kostesha¹, A. G. Laman², A. O. Shepelyakovskaya², I. S. Zaitseva¹, V. P. Orlov¹, L. A. Dykman¹, F. A. Brovko², and O. I. Sokolov^1*

¹Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, pr. Entuziastov 13, 410049 Saratov, Russia; fax: (8452) 970-383; E-mail: sokolov@ibppm.sgu.ru

²Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Institutskaya 6, 142290 Pushchino, Russia; fax: (095) 310-7007; E-mail: brovko@fibkh.serpukhov.su

^* To whom correspondence should be addressed.

Received March 30, 2004; Revision received November 17, 2004

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Single-chain miniantibodies (scFv's) to actin were obtained by the phage display method. A naive combinatorial phage display library of murine scFv's (containing 2*10⁸ independent recombinant clones) was used to select miniantibodies. After three rounds of selection two clones producing miniantibodies to chicken smooth muscle actin with affinity constants of 1.4*10⁷ and 1.2*10⁶ M^-1 were chosen. The isolated miniantibodies could specifically detect various plant and animal actins.

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KEY WORDS: actin, phage display of miniantibodies, affinity selection, single-chain miniantibodies, colloidal gold, dot-blot assay

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