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Cells were incubated for 20-30 min at 37C and then resuspended in 200 ml selection media and cultured for 3-4 days in T175 flask prior to 96 well plating. For example, by comparing similar sequences, one can identify those sections e. The plates were incubated for 1 hour at room temperature.

Next, the plates were washed using the Titertek plate washer operated using a 3-cycle wash. See, for example, Remington's Pharmaceutical Sciences, supra. In some embodiments, the antigen binding protein does not bind to the n-terminus of the catalytic domain e.

Family - As such, any comparison between these various disclosures should take this difference in numbering into account. A volume of 20-30 ml Hybridoma cells at a minimum cell density of 1×10 6 cells per ml from the 31H4 hybridoma line was placed into the small chamber of an Integra flask in Integra media see Table 5 for components of Integra media.

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Transgenic non-human animals for producing heterologous antibodies 1990-08-29 2001-10-09 Genpharm International Transgenic non-human animals for producing heterologous antibodies 1990-08-29 1998-06-23 Genpharm International, Inc. Structure 15, 545-52 2007 as the starting search model. In some embodiments, the reduction in binding is observed as a change in Bmax. In some embodiments, these antibodies bind to the variant at least twice as strongly as the wild type, and preferably 2-5,5-10, 10-100, 100-1000, 1000-10,000 fold or more to the mutant than the wild type as measured via a K d. In some embodiments, for an amino acid to be part of an epitope, the Bmax is reduced by at least 10%, for example, reductions of at least any of the following amounts: 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, or 100 percent can, in some embodiments, indicate that the residue is part of the epitope. For example, if one discovered that a change to a particular amino acid residue resulted in destroyed, undesirably reduced, or unsuitable activity, variants with such a change can be avoided. The plates were then pre-incubated for about 1 hour and 30 minutes at room temperature. In certain embodiments, amino acids in the framework regions of the variable domains are targeted. The core residues are as follows: W72, F150, A151, Q152, T214, R215, F216, H217, A220, S221, K222, S225, H226, C255, Q256, G257, K258, N317, F318, T347, L348, G349, T350, L351, E366, D367, D374, V380, S381, Q382, S383, and G384. Certain other methods for reducing immunogenicity are described, e. The results are displayed in Table 39.