Charles River Laboratories International has been granted a patent for a method of producing a population of nucleic acids encoding proteins with rabbit-derived CDR3 amino acid sequences embedded in human framework sequences. The method involves providing nucleic acids encoding rabbit CDR3 sequences and generating a population of nucleic acids encoding proteins with diversified rabbit CDR3 sequences. The human framework sequences are selected to provide a scaffold conducive for rabbit CDR3 sequences. GlobalData’s report on Charles River Laboratories International gives a 360-degree view of the company including its patenting strategy. Buy the report here.
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According to GlobalData’s company profile on Charles River Laboratories International, in-silico drug discovery was a key innovation area identified from patents. Charles River Laboratories International's grant share as of June 2023 was 1%. Grant share is based on the ratio of number of grants to total number of patents.
Method for producing nucleic acids encoding rabbit-human hybrid proteins
A recently granted patent (Publication Number: US11685773B2) describes a method for producing a population of nucleic acids that encode proteins with immunoglobulin variable domains. These proteins contain a rabbit complementarity determining region 3 (CDR3) amino acid sequence embedded in human framework sequences. The method involves providing at least 10 nucleic acids encoding rabbit CDR3 amino acid sequences and generating a population of 20 or more nucleic acids encoding proteins with rabbit CDR3 sequences embedded in human framework sequences. The nucleic acid sequences encoding the CDR1 and CDR2 amino acid sequences of the variable domain are based on either human or rabbit sequences, with modifications to incorporate amino acids from the other species. At least 80% of the nucleic acids in the population encode different CDR1 and CDR2 amino acid sequences. The human framework regions are selected to provide a scaffold conducive for rabbit CDR3 amino acid sequences, ensuring correct folding of the resulting antibodies.
In one embodiment of the method, a population of Acceptor Framework nucleic acid sequences is provided, which includes nucleic acid sequences encoding human framework regions interspaced by CDR1 and CDR2. The rabbit CDR3 amino acid sequences are then combined with the Acceptor Framework nucleic acid sequences to generate proteins with rabbit CDR3 sequences interspaced by human framework regions.
The patent also describes obtaining the rabbit CDR3 amino acid sequences from a rabbit B cell that has been immunized against an antigen of interest. Alternatively, the sequences can be determined from a sample obtained from an immunized rabbit, and lineage trees or groups of CDR3 sequences can be generated based on sequence similarities. Candidate lineages or groups can be ranked based on various factors, and an individual representative sequence can be selected to generate a nucleic acid encoding a peptide with the rabbit CDR3 amino acid sequence.
The method also includes optional modifications to the human framework sequences, such as incorporating non-human amino acids at the C-terminal ends of the human FR2 and FR3 regions. The nucleic acids encoding the rabbit CDR3 amino acid sequences can also include recognition sites for restriction enzymes, which can be used in the generation of the final nucleic acid sequences encoding the proteins.
Overall, this patent provides a method for producing a diverse population of nucleic acids encoding proteins with rabbit CDR3 sequences embedded in human framework sequences. The method allows for the generation of antibodies with desired properties and has potential applications in the field of immunotherapy and antibody engineering.
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