US 5,635,354 A · Method for describing the repertoires of antibodies (Ab) and of T-cell receptors (TcR) of an individual's immune system

Abstract

In a method for describing repertoires of antibodies (Ab) and T cell recers (TcR) of the immune system of an individual, reverse transcription is carried out on the mRNA contained in a biological sample, separate amplifications are then carried out on the transcription product (or directly on the DNA extracted from the sample) by a PCR type method for each primer pair V, C, where V corresponds to a variable segment of the repertoire of interest and C hybridizes on the constant segment of the repertoire of interest. For each J segment of the labelled repertoire, an elongation step is carried out on each of the amplification products using a specific oligonucleotide of this segment J as primer and the amplification product as matrix. For each elongation product corresponding to a triplet (V, C)J thus obtained, the size of the different elongation products is revealed. The description of the repertoires corresponds to a VCJ triplet and to the element size for each element of the repertoire.

Claims10

Dependent claims

A method for describing the repertoires of antibodies (Ab) and of T-cell receptors (TcR) of an individual's immune system, comprising the steps of:

a) obtaining a biological sample containing mRNA encoding said antibodies and T-cell receptors which contain different variable (V) and constant (C) segments,
b) reverse transcribing the mRNA to obtain a transcription product,
c) performing separate amplifications on the transcription product to form amplification products using a DNA enzymatic amplification method and separate primer pairs V,C, for each different V and C segment of the repertoire under study,
d) performing an elongation step on each amplification product for each J segment of the repertoire to form different elongation products, using as primer an oligonucleotide specific to said J segment, said oligonucleotide comprising a label,
e) revealing signals representing the molecular size of the different elongation products for each elongation product corresponding to a triplet (V,C) J thereby obtained, wherein each signal has a particular intensity, and
f) obtaining a description of the repertoires corresponding, for each segment of the repertoires, to a VCJ triplet and to the size of the segment.

3 more elements

Dependencies are inferred from the claim text.

Method for describing the repertoires of antibodies (Ab) and of T-cell receptors (TcR) of an individual's immune system

Life of this patent

Method for describing the repertoires of antibodies (Ab) and of T-cell receptors (TcR) of an individual's immune system

Life of this patent

Abstract

Claims10