Editing Trypsin (section)

==Trypsin alternatives==
{{main|Trypsin inhibitor}}
Trypsin digestion of extra cellular matrix is a common practice in cell culture. However, this enzymatic degradation of the cells can negatively effect cell viability and surface markers, especially in stem cells. There are gentler alternatives than trypsin such as Accutase which doesn't effect surface markers such as cd14, cd117, cd49f, cd292.<ref name="pmid23481571">{{cite journal | vauthors = Quan Y, Yan Y, Wang X, Fu Q, Wang W, Wu J, Yang G, Ren J, Wang Y | title = Impact of cell dissociation on identification of breast cancer stem cells | journal = Cancer Biomarkers| volume = 12 | issue = 3 | pages = 125–33 | date = 2012 | pmid = 23481571 | doi = 10.3233/CBM-130300 }}</ref><ref name="pmid30983404">{{cite journal | vauthors = Skog M, Sivlér P, Steinvall I, Aili D, Sjöberg F, Elmasry M | title = The Effect of Enzymatic Digestion on Cultured Epithelial Autografts | journal = Cell Transplantation | volume = 28 | issue = 5 | pages = 638–644 | date = May 2019 | pmid = 30983404 | pmc = 7103596 | doi = 10.1177/0963689719833305 }}</ref> However Accutase decreases the surface levels of FasL and [[Fas receptor]] on [[macrophage]]s, these receptors are associated with cell [[cytotoxicity]] in the immune system and can also facilitate [[apoptosis]]-related cell death.<ref name="pmid35383242">{{cite journal | vauthors = Lai TY, Cao J, Ou-Yang P, Tsai CY, Lin CW, Chen CC, Tsai MK, Lee CY | title = Different methods of detaching adherent cells and their effects on the cell surface expression of Fas receptor and Fas ligand | journal = Scientific Reports | volume = 12 | issue = 1 | pages = 5713 | date = April 2022 | pmid = 35383242 | pmc = 8983651 | doi = 10.1038/s41598-022-09605-y | bibcode = 2022NatSR..12.5713L }}</ref>

ProAlanase could also serve as an alternative to Trypsin in proteomic applications.<ref name="Samodova_2020">{{cite journal | vauthors = Samodova D, Hosfield CM, Cramer CN, Giuli MV, Cappellini E, Franciosa G, Rosenblatt MM, Kelstrup CD, Olsen JV | title = ProAlanase is an Effective Alternative to Trypsin for Proteomics Applications and Disulfide Bond Mapping | journal = Molecular & Cellular Proteomics | volume = 19 | issue = 12 | pages = 2139–2157 | date = December 2020 | pmid = 33020190 | pmc = 7710147 | doi = 10.1074/mcp.tir120.002129 | doi-access = free }}</ref> ProAlanase is an ''Aspergillus niger'' fungus protease that can achieve high proteolytic activity and specificity for digestion under the correct conditions.<ref name="Samodova_2020" /> ProAnalase, the acidic prolyl-endopeptidase protease, previously studied as An-PEP, has been observed in various experiments to define its specificity.<ref name="Samodova_2020" /> ProAnalase performed optimally in LC-MS applications with short digestion times and highly acidic pH.<ref name="Samodova_2020" />