Method for cloning cognate antibodies

C - Chemistry – Metallurgy – 12 – N

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C12N 15/10 (2006.01) C07K 16/00 (2006.01)

Patent

CA 2677771

The invention relates to a procedure for linking cognate pairs of VH and VL encoding sequences from a population of cells enriched in particular surface antigen markers. The linking procedure involves a multiplex molecular amplification procedure capable of linking nucleotide sequences of interest in connection with the amplification, in particular polymerase chain reaction (multiplex PCR). The method is particularly advantageous for the generation of cognate pair libraries as well as combinatorial libraries of variable region encoding sequences from immunoglobulins. The invention also relates to methods for generation of chimeric human/non-human antibodies and expression libraries generated by such methods.

La présente invention concerne une procédure de liaison de paires apparentées de séquences codant VH et VL prises dans une population de cellules enrichies en marqueurs particuliers d'antigènes de surface. La procédure de liaison implique une procédure d'amplification moléculaire multiplex capable de lier les séquences de nucléotides étudiées en relation avec l'amplification, et notamment la PCR multiplex. Ce procédé donne de bons résultats surtout pour la génération d'échantillothèque de paires apparentées, ainsi que d'échantillothèques combinatoires de séquence codant des régions variables des immunoglobulines. L'invention concerne également des procédés de génération d'anticorps chimériques humains/non-humains et les échantillothèques d'expressions produites selon ces procédés.

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