Secretory expression of human ScFv against keratin in Pichia pastoris

Objective:To express secretively human anti-keratin ScFv in Pichia pastoris.Methods:Anti-keratin ScFv gene from plasmid p3MH/ScFv was subcloned into vector pPIC9K.After confirmed by DNA sequence analysis,the recombinant plasmid pPIC9K/ScFv was transducted into the genome of GS115 Pichia pastoris.Mut~s mutiple insert transformants were screened by G418 and induced by 5 ml/L methanol to express soluble ScFv.Results:After 6 days of methanol induction,anti-keratin ScFv was efficiently secreted into the medium.Western blot proved that the expressed protein had specific keratin binding activity.Conclusion:The successful expression of anti-keratin ScFv in Pichia pastoris laid a solid foundation for its further application.