Monoclonal antibody production in b cells and uses therof

A method for increasing efficiency of EBV transformation of primary B cells, the method comprising:

• (i) providing a primary B cell;
• and
• (ii) transforming the primary B cell using EBV in the presence of an inhibitor of a host innate response to EBV-mediated proliferative signals,
• wherein the efficiency of the EBV transformation of the primary B cell is increased compared to EBV transformation without the use of an inhibitor of a host innate response to EBV-mediated proliferative signals.

A method for producing a clone of an immortalized primary B cell capable of producing a monoclonal antibody with a desired antigen specificity, the method comprising:

• (i) transforming a population of cells comprising primary B cells with EBV in the presence of an inhibitor of a host innate response to EBV-mediated proliferative signals;
• (ii) screening the supernatant from the population of cells for antigen specificity;
• and
• (iii) isolating from the population of cells an immortalized B cell clone capable of producing a monoclonal antibody having the desired antigen specificity.

A method for producing a monoclonal antibody, comprising the steps of:

• (i) providing to a primary B cell in culture an activator of B cell proliferation in the presence of an inhibitor of a host innate response to activator-mediated proliferative signals;
• (ii) screening the supernatant from the culture for antigen specificity;
• (iii) isolating from the culture a B cell capable of producing a monoclonal antibody having the desired antigen specificity;
• and
• (iv) expressing the monoclonal antibody, comprising
• (a) culturing the B cell under conditions where the monoclonal antibody is expressed;
• or
• (b) obtaining a nucleic acid encoding the antibody of interest from the isolated B cell;
• (c) employing the nucleic acid to prepare an expression host that can express the antibody of interest;
• and
• (d) culturing the expression host under conditions where the antibody of interest is expressed.

A method for identifying a novel broadly neutralizing antibody having a desired antigen specificity, comprising the steps of:

• (i) providing a subject infected with or possessing the antigen for which the antibody specificity is desired;
• (ii) culturing primary B cells obtained from the subject;
• (iii) providing to the cultured primary B cells an activator of B cell proliferation in the presence of an inhibitor of a host innate response to activator-mediated proliferative signals;
• and
• (iv) screening the supernatant from the culture of proliferating primary B cells for a novel broadly neutralizing antibody having the desired antigen specificity.

A method for generating a profile of the humoral response of a subject, comprising:

• (i) culturing primary B cells obtained from a subject;
• (ii) providing to the cultured primary B cells an activator of B cell proliferation in the presence of an inhibitor of a host innate response to activator-mediated proliferative signals;
• (iii) screening the supernatant from the culture of proliferating primary B cells for one or more antibodies;
• and
• (iv) characterizing the one or more antibodies, wherein a profile of the humoral response of the subject is generated.