Сибирский федеральный университет

Тип публикации: статья из журнала

Год издания: 2002

Идентификатор DOI: 10.1023/A:1015065323342

Ключевые слова: affinity selection; antibody phage display library; granulocyte colony-stimulating factor; murine single-chain Fv

Аннотация: To develop a phage display of single-chain antibodies (scFv), fractions of total cell DNA and RNA were obtained from splenocytes of naive mice. The DNA fragments encoding variable regions of light and heavy immunoglobulin chains were amplified and isolated using primers specific to the conservative regions of these genes. The construction of the library was based on the principle of stochastic combining the DNA fragments encoding the light and heavy antibody chains with the DNA linker, whose structure corresponded to the (GIy(4)Ser)(3) sequence. The scFv library was constructed using the E. coli TG1 strain and the phagemid vector pHEN1. The repertoire of the library exceeded 5 x 10(7) independent recombinant clones. The clones producing antibodies to human granulocyte colony-stimulating factor were isolated. The affinity constants of the resulting scFv were in the range of 2 x 10(4) to 1.8 x 107 M-1.

Журнал: RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY

Выпуск журнала: Vol. 28, Is. 2

Номера страниц: 108-115

ISSN журнала: 10681620

Место издания: NEW YORK

Издатель: MAIK NAUKA/INTERPERIODICA

• Laman A.G. (KRASNOYARSK STATE UNIV)
• Shepelyakovskaya A.O. (KRASNOYARSK STATE UNIV)
• Ulitin A.B. (KRASNOYARSK STATE UNIV)
• Markova E.V. (KRASNOYARSK STATE UNIV)
• Mareeva T.Y. (KRASNOYARSK STATE UNIV)
• Bystrov N.S. (KRASNOYARSK STATE UNIV)
• Brovko F.A. (KRASNOYARSK STATE UNIV)
• Nesmeyanov V.A. (KRASNOYARSK STATE UNIV)

• Web of Science Core Collection