Compositions and methods to modulate cell membrane resealing
A technology of composition and cell membrane, applied in the field of polypeptide composition regulating cell membrane repair, can solve the problem that the nucleation process has not been completely solved
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[0213] The preparation of monoclonal antibodies is well known in the art; see for example (Harlow et al., Antibodies: A Laboratory Manual, page 726 (Cold Spring Harbor Pub. 1988). Monoclonal antibodies can be prepared by injection with a composition comprising the antigen Mice or rabbits, serum samples taken to verify the presence of antibody production, spleen removed to obtain B lymphocytes, lymphocytes fused with myeloma cells to create hybridomas, hybridomas cloned, selection of clones that produce antibodies against antigens , and by isolating antibodies from hybridoma cultures. Monoclonal antibodies can be isolated and purified from hybridoma cultures by techniques well known in the art.
[0214] In other embodiments, antibodies can be produced recombinantly, such as by phage display or by combinatorial methods. Phage display and combinatorial methods can be used to isolate recombinant antibodies that bind to membrane repair polypeptides, MG53, membrane repair polypeptid...
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[0239] Discovery of MG53, a muscle-specific TRIM family protein . MG53 was isolated using a previously established immunoproteomic approach that allowed the identification of proteins associated with myogenesis, Ca 2- Novel proteins involved in signaling and maintaining membrane integrity. Briefly, the method employs a monoclonal antibody library comprising ~6500 clones generated from mice immunized with triplet-rich membranes from rabbit skeletal muscle. Antibodies of interest were selected based on the z-line staining pattern of striated muscle sections observed under immunofluorescence microscopy. The target protein was purified by antibody affinity column, and the partial amino acid sequence of the purified protein was obtained. Based on the partial amino acid sequence, the complete cDNA encoding the target gene can be isolated from the skeletal muscle cDNA library. Homologous gene screening was then employed to search for the presence of different isoforms of the ide...
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