Construction Of A Phage Display Library Of Single-chain Fv Antibodies Repertoire From Recombinant Human Granulocyte Colony-Stimulation Factor (G-CSF) Immunized Mouse And Identification Of Anti-G-CSF Antibody Fragments And V-region Genes.

Granulocyte colony-stimulating facter (G-CSF) is a hematopoietic cytokine produced by monocytes fibroblasts and endothelial cells. G-CSF is known to have multiple functions in normal steady-state hematopoiesis such as the regulation of neutrophil production and release from the bone marrow, neutrophil progenitor proliferation and differentiation and the state of functional activation of neutrophils. But the serious autodamage of inflammatory and other serious pathological process induced by superoxide free residue with PMN (neutrophi) respiratory burst metabolsm are relational with the high level of G-CSF in peripheral circulation. So it is important to keep the kinetic balance of neutrophil in human body and the usage of anti-G-CSF antibody is the one of the methods.Phage display technology of immunoglobulin repertoire is a main outbreakof antibody engineering. We use this technology to construct a phage display library of immunoglobulin repertoire from human G-CSF immunized mouse and i-dentify the generated anti G-CSF antibody fragment as well as their V-region genes. The heavy-chain and kappa light-chain variable region gene repertoire of immunoglobulin were separately amplified from the spleen cell mRNA of a human G-CSF immunized mouse by RT-PCR and joined by a DNA linker encoding peptide (GGGGS)₃ as a single-chain Fv (ScFv) DNA frangment with overlap ex-tensien. The ScFv DNA fragment was cloned into a phagemid (pCANTAB 5E)...