Escherichia coli thermosensitive enterotoxin activity and stability detection and preparation of polyclonal antibody

• Main Authors: SHI XIAOJIE, ZHANG QIAN, MA WEICHAO, LU GUANJIE
• Format: Patent
• Language: Chinese; English
• Published: 23.12.2022
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; TESTING; VINEGAR; WINE

The invention discloses escherichia coli thermosensitive enterotoxin activity and stability detection and polyclonal antibody preparation, which comprises the following steps: toxin in-vitro activity verification: detecting purified LT protein concentration by using a Bradford kit, and diluting the LT protein concentration to 100 [mu] g.mL <-1 >, 10 [mu] g.mL <-1 >, 1 [mu] g.mL <-1 >, 0.1 [mu] g.mL <-1 > and 0.01 [mu] g.mL <-1 > by using a complete medium; the method comprises the following steps: resuscitating Vero cells for subculture, counting digested cells, inoculating the cells into a 24-pore plate, and culturing at 37 DEG C in 5% CO2; setting three repetitions in each of an LT protein treatment group and a blank control group after cell adherence; after culturing for 3 hours, photographing and observing cell morphological change. The extracted LT protein is good in activity, and the freeze-dried LT protein is optimal in preservation stability at 4 DEG C; the polyclonal antibody prepared after multiple