Method of obtaining antibodies of interest and nucleotides encoding same

摘要

The invention is a methodology which makes it possible to select from a very large number of cells, a single cell or cells of interest and obtain specific information from those cells in a rapid and efficient manner. As an example of the methodology, a large number of antibody producing cells such as plasma cells are separated so that these individual antibody producing plasma cells are placed in individual wells. The cells are allowed to produce antibodies and the antibodies in the wells are then contacted with a protein bound to a solid surface such as a well top. The protein universally and specifically binds antibodies in the wells. The surface or well tray top includes addresses configured such that each address is specifically related to one of the individual wells containing a cell producing antibodies.

主权项

1. A method of obtaining antigen binding and nucleic acid sequence information about antibodies produced from a plurality of isolated antibody producing cells, comprising the steps of:
placing a plurality of individual antibody producing cells in a plurality of individual wells; contacting antibodies produced by the antibody producing cells in the wells with an antibody binding agent bound to a removable first top, to obtain surface bound antibodies, wherein the first removable top comprises regions relatable to the individual wells; simultaneously determining binding information relating to binding of the antibodies bound to the first removable top to a known antigen and associating the binding information to a particular region of the first removable top and an individual well; lysing the antibody-producing cells in the wells; hybridizing mRNAs from the lysed antibody-producing cells with oligonucleotide probes comprising 1) tags, and 2) antibody light or heavy chain nucleic acid sequences that selectively bind to mRNAs that encode antibodies, wherein the oligonucleotide probes are bound to a second removable top, wherein the second removable top comprises regions relatable to the individual wells, and wherein the tags in each region are different; converting the mRNAs hybridized to the second removable top into cDNAs which comprise the tags specific to the regions on the second surface; releasing the tagged cDNAs from the second top; pooling the tagged cDNAs; sequencing the tagged cDNAs; and utilizing the tags to associate the cDNA sequences to a particular well, which associates the cDNA sequences with binding information about the antibody produced by the antibody-producing cells in the wells.