Lipopolysaccharide conjugated protein and monoclonal antibody and usage

Abstract

The present invention discloses one kind of lipopolysaccharide bindin and its monoclonal antibody, preparation process and use. Through extracting NH-LBP coding cDNA segment from human liver cell, preparing NH-LBP protein with insect sf21 cell, establishing humanized phage antibody base with pComb3H phagemid and screening with NH-LBP as antigen, anti-NH-LBP humanized monoclonal antibodies, Fab antibody and dsFv antibody are prepared. The Fab antibody and the dsFv antibody can combine specifically with the N end of complete human lipopolysaccharide bindin and inhibit the combination between LBP and LPS competively to lower the LPS sensitivity of body, lower LPS caused excessive inflammatory reaction, promote the degradation of LPS in body and prevent diffused blood coagulation inside capillary vessel and multiple organ failure.

Description

technical field [0001] The invention relates to genetic recombination engineering, in particular to lipopolysaccharide-binding protein and its monoclonal antibody, and the application of lipopolysaccharide-binding protein and its monoclonal antibody. Background technique [0002] Lipopolysaccharide binding protein (LBP) is mainly produced by hepatocytes (renal proximal tubule epithelial cells, alveolar wall type II cells can also be synthesized in a small amount) and mediates the binding of endotoxin to target cell receptors. Acute-phase stress response protein, which can significantly promote the combination of lipopolysaccharide (LPS) and CD14 (mCD14) on the surface of monocyte / macrophage membrane to form LPS-LBP-CD14 complex, which passes through the membrane of monocyte / macrophage The Toll-Like Receptor 4 (TLR4)-Myeloid Differentiation Protein 2 (MD-2)-MYD88 recognition complex is transduced, and then transduced into the nucleus by polydominant transcription nuclear fact...

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Problems solved by technology

[0004] Lipopolysaccharide binding protein can significantly promote the combination of lipopolysaccharide and mCD14. Nanbo et al. found that human lipopolysaccharide binding protein can increase the sensitivity of the body to lipopolysaccharide recognition by 1000 times. In the early stage of acute inflammation reaction, when the serum lipopolysaccharide concentration is relatively high When it is low, there is already a small amount of lipopolysaccharide-binding protein involved, and only in the presence of lipopolysaccharide-binding protein can the body respond to a small dose of lipopolysaccharide or bacteria, thereby enhancing the body's anti-infection effect, but in the middle and late stages of the acute inflammatory response , when the serum lipopolysaccharide-binding protein content rises, lipopolysaccharide-binding protein will lead to excessive activation of monocytes / macrophages, and a large amount of secretion of interleukin (IL)-1β, IL-6, tumor necrosis factor-α (TNF- α) and other cytokines, these factors strongly accelerate the transcription of lipopolysaccharide-binding protein gene and the synthesis of lipopolysaccharide-binding protein polypeptide, which will lead to serious damage to the body, septic shock, disseminated intracapillary coagulation (DIC), multiple Organ failure (MOF) (see Eur J Biochem 1999, 260 (1): 183-191), etc., so how to suppress excessive inflammatory response and prevent the occurrence of disseminated intracapillary coagulation and multiple organ failure has become the current clinical practice. An important research direction in the work

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