Methods and compositions for generating human monoclonal antibodies

An isolated population of non-transformed human B cell clones that produce human monoclonal antibodies immunoreactive with cell surface antigens, wherein said isolated population of non-transformed human B cell clones is generated by:

• (a) (1) culturing in vitro in an immunization medium, an isolated population of human lymphocytes comprising a plurality of non-transformed human B lymphocytes and cells that natively express said cell surface antigens that have been immobilized to a solid substrate, and thereby contacting said isolated population of human lymphocytes comprising said plurality of non-transformed human B lymphocytes with said cells that natively express said cell surface antigens that have been immobilized to the solid substrate, and
• (2) allowing one or more non-transformed human B lymphocytes to specifically bind to the immobilized cells bearing said cell surface antigens,
• wherein the surfaces of the non-transformed human B lymphocytes are free of serum, and
• (b) removing unbound B lymphocytes thereby selecting the non-transformed human B lymphocytes that specifically bind to the cells bearing the cell surface antigens, and
• (c) culturing the selected B lymphocytes of (b) under conditions favorable for B cell proliferation to yield a plurality of non-transformed B cell clones that produce human monoclonal antibodies immunoreactive with the cell surface antigens.