RESUMO
Objective To prepare a specific anti-lactoferrin single chain variable fragment(ScFv).Methods Anti-lactoferrin clones were screened from a 'naive' phage antibody library against the immobilized lactoferrin antigen,then the clones were transformed to the E.coli HB2151 to give soluble expression of antibody fragments.The culture supernatant containing ScFv was purified by immobilized metal affinity chromatography,and then determined with SDS-PAGE and ELISA.Results The results demonstrated that ScFvs were specific;they did not react with transferrin,lysozyme and bovine serum albumin in ELISA.The SDS-PAGE showed that the ScFvs had high purity through affinity chromatography and the molecular weight of them was about 32 kD.Conclusions The successful generation of the ScFvs against lactoferrin provides a basis for further study and clinical applications for dry eye and other ocular diseases.
RESUMO
Triptophenolide (TN) is a colorless crystalline plate isolated from ethyl acetate extracts of Tripterygium wilfordii. It can obviously inhibit the edema caused by xylol and mixture croton oil; and decrease the concentration of Vitamin C in the adrenal gland of mice.TN can remarkably inhibit the delayed type hypersensitivity (DTH) reaction induced by DNCB and BSA; and diminished the peripheral blood ANAE+lymphocytes in rats and micc.Moreover, TN can dramatically increase the amount of total serum complement and significautly decrcase the serum antibody products (1gG ) of rats and mice. The phagocytosis of perioneal exudate macrophages in mice present double effects in vitro. TN can promote the phagocytosis of macrophages at the small dosage, but at larger dosage, it showed inhibition.The results mentioned above showed that triptophenolide may be one of the main effective component of T.wilfordii for treating auto-immune diseases.
