Methods for Screening B Cell Lymphocytes

A method of detecting a B cell lymphocyte expressing an antibody that specifically binds to an antigen of interest for preparation of a cDNA library, the method comprising:

• introducing a sample comprising B cell lymphocytes into a microfluidic device, the microfluidic device comprising:
• an enclosure having a flow region and a sequestration pen, wherein said sequestration pen comprises an isolation region having a single opening and a connection region, said connection region providing a fluidic connection between said isolation region and said flow region, and wherein said isolation region of said sequestration pen is an unswept region of said microfluidic device;
• loading a B cell lymphocyte from said sample into said isolation region of said sequestration pen;
• introducing said antigen of interest into said flow region of said enclosure such that said antigen of interest is proximal to said B cell lymphocyte;
• and
• monitoring binding of said antigen of interest to said antibody expressed by said B cell lymphocyte disposed on a surface of said isolation region, wherein said isolation region of said sequestration pen comprises at least one conditioned surface, thereby detecting an antibody-expressing B cell lymphocyte of interest;
• and lysing the B cell lymphocyte of interest, thereby releasing RNA.