ABSTRACT
The use of immune checkpoint inhibitors (ICIs) has changed the clinical outcome of non-small cell lung cancer (NSCLC), with the widespread application of ICIs, immune-related adverse events (irAEs) have also appeared. Immune checkpoint inhibitor pneumonitis (CIP) is a serious adverse event of ICIs treatment that needs attention. Therefore, early identification of high-risk groups of CIPs and early intervention can reduce the occurrence of permanent drug withdrawal and severe CIPs, thereby improving patients′ prognosis.
ABSTRACT
Objective To prepare the mouse anti recombinant human Galectin-7 antibody and the antibody was characterized in bladder cancer.Methods The gene coding for Galectin-7 was amplified by PCR from the cDNA of human foreskin cells and cloned into prokaryotic expression vector pET28a.Then the recombinant plasmid pET28a/Galectin-7 was transformed into E.coli BL21 (DE3)and expressed under IPTG induction.The recombinant Galectin-7 was purified through Ni2+-NT agarosegel column and the purified Galectin-7 used as imunogen to imunize the mouse.The titer and specificity of the anti-Galectin-7antibody from the mouse were analyzed by ELISA,Western blot and immunohistochemistry,respectively.Results The recombinant Galectin-7 was successfully expressed and purified,and the polyclonal ani-Galectin-7 antibody was suc-cessfully prepared.The titer of the antiserum was 1∶32 000 by ELISA.Western blot analysis showed this antiboday reacted specifically with Galectin-7.Immunohistochemistry analysis showed the antibody could recognize the native Galectin-7 in the human bladder cancer tissue.Conclusion The preparation recombinant Galectin-7 protein was as immunogen in rabbits.It was successful to produce high titer and high specificity of anti Galectin-7 polycolonal antibody.