Profiling of adaptive immunity and methods for generating monoclonal antibodies
2010
A method of generating monoclonal antibodies, comprising the steps of: a) creating a library of cDNA sequences, the creation comprising the steps of: (i) isolating mRNA isolated lymphocytes from a host subject has been immunized with a antigen; (Ii) reverse transcribe mRNA into cDNA; (Iii) amplifying the cDNA sequences of the target antibody; and (iv) sequencing the cDNA sequences of the target antibody; b) analyze the frequency of the cDNA sequences of the target antibody, comprising: (i) compare the relative frequencies of the cDNA sequences of the target antibody before and after exposure to the antigen, where the cDNA sequences of the target antibody which they have a significantly higher frequency after antigen challenge are identified as candidate antibody sequences; or (ii) compare the relative frequencies of the cDNA sequences of the target antibody in two or more times after exposure to the antigen, where the cDNA sequences of the target antibody having a significantly lower frequency at later times are identified as sequences antibody candidates.
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