Method for preparing tumor-associated antigen NY-ESO-1 by taking pGEX6P1 as carrier and application of tumor-associated antigen NY-ESO-1

• Main Authors: HE XIAOBO, ZHANG YA'NAN, DING KAI, WU YANFENG, YU GANJUN
• Format: Patent
• Language: Chinese; English
• Published: 15.09.2023
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; HUMAN NECESSITIES; HYGIENE; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEDICAL OR VETERINARY SCIENCE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PHYSICS; PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS ORMEDICINAL PREPARATIONS; SPIRITS; TESTING; VINEGAR; WINE

The invention relates to the technical field of biological medicines, and provides a method for preparing a tumor-associated antigen NY-ESO-1 by taking pGEX6P1 as a carrier and application of the tumor-associated antigen NY-ESO-1. The method comprises the following steps: recombining an NY-ESO-1 gene and a pGEX6P1 vector with GST tag protein, transfecting competent cells, sequentially carrying out seed acquisition and amplification culture, carrying out IPTG induction, breaking bacteria, collecting supernatant, and purifying to obtain the mg-level GST/NY-ESO-1 fusion protein. According to the invention, the pGEX6P1 vector is selected as a bearing vector, and various technological processes and conditions are optimized in the subsequent expression and purification process, so that the target protein appears in the soluble supernatant, the step of renaturation of the inclusion body in the existing inclusion body preparation mode is avoided, and the separation and purification process and the operation difficult