METHODS FOR DETERMINING LYMPHOCYTE RECEPTOR CHAIN PAIRS

摘要

Provided herein are high-throughput sequencing methods to study the diversity and functionality of lymphocyte receptor chains and pairing of the same. Specifically, the methods provided herein are used to identify with confidence one or more lymphocyte receptor chain pairs in a sample, for example one or more functional chain pairs.

主权项

1. A method for identifying a plurality of lymphocyte receptor chain pairs in a sample comprising a plurality of lymphocytes or progeny thereof, wherein each of the lymphocyte receptor chain pairs is from a single lymphocyte clone, comprising,
subjecting the sample to conditions suitable for expansion of one or more of the lymphocytes or progeny thereof to form an expanded sample; partitioning the expanded sample into a plurality of individual vessels to provide a plurality of sample subpopulations; optionally subjecting one or more of the sample subpopulations to conditions suitable for expansion of one or more of the lymphocytes or progeny thereof in the one or more of the sample subpopulations; to optionally form one or more expanded sample subpopulations; sequencing nucleic acid clonotypes encoding the lymphocyte receptor chains in each sample subpopulation or expanded sample subpopulation to determine the identity of the lymphocyte receptor chains in each sample subpopulation or expanded sample subpopulations; determining the observed distribution of each of the lymphocyte receptor chains across the subpopulations and calculating statistical probabilities that the observed distributions of lymphocyte receptor chain pairs in the subpopulations are independent from one another; and identifying the plurality of lymphocyte receptor chain pairs based on the calculated statistical probabilities.