EBC product recommendations | Recombinant Trypsin Digestion — a better choice for cell dissociation

Trypsin is a serine protease that specifically cleaves lysine and arginine C-terminal peptide bonds. Traditional trypsin is extracted from the pancreas of animals such as pigs, cattle and sheep, and has been widely used in the production of biological products. Animal-derived trypsin is at risk of introducing exogenous viruses, for example, in the 1960s, there were incidents in which SV40 virus and avian leukemia virus were detected in polio vaccines, attenuated yellow fever live vaccines, and in 2010, a manufacturer’s rotavirus vaccine detected a porcine circovirus sequence. The third part of the 2020 edition of the Chinese Pharmacopoeia states: “The raw materials and excipients of biological products … Raw materials and excipients with no virus safety risk or low risk should be selected as much as possible for the production of biological products, such as the use of recombinant technology to replace animal-derived biological materials” (Chinese Pharmacopoeia 2020 Edition, Part III, 25 pages).

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Trypsin molecular structure

The recombinant trypsin expressed by E. coli has a similar effect to trypsin extracted from the animal pancreas, and after purification consists of no less than 70% β-Trypsin and no more than 20% α-Trypsin.

Hzymes recombinant trypsin digestion solution is a self-developed high-purity recombinant enzyme digestion produced by microbial fermentation, used to dissociate a variety of adherent mammalian cells, including VERO, MDCK, human diploid cells, 293, CHO-K1, chicken embryonic cells, etc., can be widely used in a variety of biotechnology processes: such as the acquisition of primary cells, digestion dissociation of tissue blocks, passage digestion of adherent cells, specific proteolysis, etc. It is more specific and less toxic.

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Recombinant trypsin digestion has several advantages over animal-derived trypsin:

Mild action—Only one enzyme action reduces dissociation of multiple proteases in trypsin and other extraction reagents to maintain cell health

Stable activity — 2-8° storage and transport, no -20° storage required

Easy to use – implanted directly into experiments and production protocols

Non-animal source – no exogenous virus contamination, no need to detect the virus, in line with the national pharmacopoeia for biological products virus safety control requirements

Aseptic packaging – safer to use

Domestic original research – cost-effective, supply guarantee

Indicators of recombinant trypsin digestion

Definition of enzyme activity: 25 °C, pH 7.6, reaction system 3.2 ml (1 cm optical path), enzymatic digestion of BAEE per minute increases the absorption value at 253 nm by 0.003 defined as one trypsin unit (USP)

Freeze-thaw stability: No loss of enzyme activity was observed after 6 freeze-thaws in 4 different batches

Freeze-thaw stability:

Temperature stability: Recombinant trypsin digestion solution did not decrease in activity for 8 weeks at 4 °C or room temperature protected from light

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Enzyme purity: Unlike trypsin expressed by animal sources and E. coli, chromatography shows as a single enzyme.

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Short-term toxicity: MDXK cells after 270 minutes of exposure to recombinant trypsin digestion solution, cell viability did not decrease.

hzymes Varible

A variety of specifications for your application

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