Method for proteomic analysis utilizing immune recognition and cumulative subtraction

摘要

A method of identifying proteins in a proteome comprises the steps of: (a) immunizing a host organism with a sample containing an initial mixture of proteins from a tissue or fluid to be evaluated to elicit an antibody response to one or more proteins in the initial mixture; (b) isolating an antiserum from the host organism after immunization; (c) contacting the antiserum with an array of known proteins to form one or more antibody-protein conjugates, thereby identifying the one or more proteins in the initial mixture that elicited an antibody response in the host organism; (d) optionally removing the proteins that elicited antibody formation in step (a) from the initial mixture to form a mixture of non-responding proteins; and (e) optionally repeating the previous steps (a) through (d) one or more times, each repetition utilizing the mixture of non-responding proteins from the immediate prior repetition in place of the initial mixture of proteins to identify one or more additional proteins present in the tissue or fluid to be evaluated. A corresponding method of preparing antisera for identifying proteins in a proteome is also disclosed.