The DMSO is essential as it minimizes the size of water crystals formed. Lymphocytes are stored in a solution containing 90% fetal calf serum (FCS) and 10% dimethylsulfoxide (DMSO). In addition, it may affect assays involving oxidative enzymes. This stops bacterial growth but has little effect on proteolytic activity. 3.Ĭhemicals such as sodium azide or sodium fluoride can be added. 2.īefore or after heating, the sample can be frozen (in aliquots) at −70☌. This destroys most but not all proteolytic activity. The sample can be heated to 60☌ for 5–10 min with little loss in antibody binding capacity. ![]() This can be circumvented by employing the following procedures: 1. However, antibodies obtained as serum (polyclonal) or as tissue culture supernatant (monoclonal) are contaminated with proteases and serum samples are generally collected under conditions that render them nonsterile. As a pure, sterile solution, they will maintain activity for several weeks at room temperature and several months at 40☌. IgG antibodies are very stable molecules held together by several disulfide bonds. 81 However, the ADCC enhancement is observed with alteration to the nonfucosylated hybrid type is not beyond that to the core nonfucosylated complex-type Fc oligosaccharide. 93 It has been demonstrated that the ADCC activity of antibody IgG's with nonfucosylated hybrid-type Fc oligosaccharides is significantly enhanced through the FcγRIIIa, as compared to the ADCC activity of the native unmodified core-fucosylated complex-type IgG's produced by the same parent mouse hybridoma. 81 Although the precise mechanism for altering the oligosaccharide structure of glycoproteins by monensin is not yet clear, it is known that glycoproteins with immature oligosaccharide moieties are secreted (reaching the cell surface and released into the culture medium), most probably due to the monensin-induced inhibition of intracellular transport between Golgi apparatuses. Hanai, in Comprehensive Glycoscience, 2007 4.33.3.3.2.2 Nonfucosylated hybrid-type glycoformsĪntibody IgG's bearing Fc oligosaccharides of the nonfucosylated hybrid type were first generated by mouse hybridoma cultures in the presence of monensin, an ionophore that strongly binds the primary ion, and the biological activity of these IgG's was evaluated.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |