US 8,273,688 B2Granted

Collection of HCDR3 regions and uses therefor

AssigneeMorphosys AG

InventorsMarkus Enzelberger, Stefanie Thiel, Josef Prassler, Stefanie Urlinger, Christine Rothe

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Abstract

The present invention is directed to the preparation and use of a collection of antibody heavy chain complementarity determining region 3 (HCDR3) members, where diversity of the collection is a function of the length of the HCDR3 members. The diversity of the collection of HCDR3 regions substantially represents the natural amino acid distribution of HCDR3 in the human repertoire. This natural amino acid distribution can be represented by biasing the complete random distribution of amino acids, accordingly, in the HCDR3 encoding DNA sequence by using trinucleotide mutagenesis (TRIM) technology. A collection of HCDR3 members of the invention each can be comprised within a variable region of an antibody (or fragment thereof) to form a library of synthetic antibodies or antibody fragments. The invention also provides nucleic acid molecules encoding such diverse collection and methods of making and using the same.

Claims7

Dependent claims

A collection comprising diverse, isolated human or humanized antibody H-CDR3 regions, wherein the H-CDR3 regions comprise a first, second and third range of amino acid lengths, wherein the first range consists of 4 to 7 amino acids in length, wherein the amino acids of the first range of lengths comprise diversity according to a first diversity factor, wherein the second range consists of 8 to 14 amino acids in length, wherein the amino acids of the second range of lengths comprise diversity according to a second diversity factor, and wherein the third range consists of 15 to 23 amino acids in length, wherein the amino acids of the third range of lengths comprise diversity according to a third diversity factor, wherein the diversity factors comprise a frequency rate at which amino acids appear at each position in the H-CDR3 region, and wherein the first, second and third diversity factors are different, wherein the H-CDR3 regions consisting of a length of 4 amino acids comprise at position 1 Aspartic acid and Glycine have a frequency rate of about 25, Valine, Glutamic acid, Alanine, Leucine, Arginine and Serine have a frequency rate of about 5%, all other amino acids except Cysteine have a frequency rate of about 1.1%;

at position 2 Arginine, Glycine, Serine and Tyrosine have a frequency rate of about 10%, Alanine, Aspartic acid, Histidine, Isoleucine and Threonine have a frequency rate of about 5% and all other amino acids except Cysteine have a frequency rate of about 1.8%;
at position 3 Glycine has a frequency rate of about 25%, Tyrosine, Serine, Aspartic Acid and Arginine have a frequency rate of about 10% and all other amino acids except Cysteine have a frequency rate of about 1.8%;
and at position 4 Glycine has a frequency rate of about 25%, Tyrosine, Serine, and Alanine have a frequency rate of about 10% and all other amino acids except Cysteine have a frequency rate of about 2.4%.

Dependencies are inferred from the claim text.