BioAtla has been granted a patent for a method of preparing a conditionally active polypeptide or antibody. The method involves evolving the DNA encoding the parent polypeptide or antibody to increase the net charge or number of acidic amino acid residues, expressing the mutant DNA to obtain mutant polypeptides or antibodies, and selecting the conditionally active variant based on its decreased activity under specific conditions. The patent also covers pharmaceutical compositions, nanoparticle and drug conjugates, and various uses of the conditionally active polypeptide or antibody. GlobalData’s report on BioAtla gives a 360-degree view of the company including its patenting strategy. Buy the report here.
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Method of preparing conditionally active antibodies or antibody fragments
A recently granted patent (Publication Number: US11773509B2) describes a method for preparing a conditionally active antibody or antibody fragment. The method involves evolving a DNA encoding the parent antibody, single chain antibody, or antibody fragment to increase the number of acidic amino acid residues, creating mutant DNAs. These mutant DNAs are then expressed to obtain mutant antibodies or antibody fragments. The conditionally active antibody or antibody fragment is selected from the mutant antibodies or antibody fragments based on its reversible inactivation in a first assay at a pH of 7.4 and a decrease in activity compared to a second assay at a pH of 6.0.
The evolving step in the method can employ site-directed mutagenesis techniques such as oligonucleotide-mediated mutagenesis, PCR mutagenesis, or cassette mutagenesis. It can also involve introducing one or more codons of acidic amino acids into the DNA encoding the parent antibody, single chain antibody, or antibody fragment. This can be done through codon substitution, codon insertion, or both.
The introduced codons can be placed in different regions of the DNA encoding the parent antibody, single chain antibody, or antibody fragment. They can be inserted into the region that encodes the active site of the antibody or into a region outside of the active site. Additionally, the DNA can be evolved to introduce codons of acidic amino acid residues into the complementarity determining regions of the antibody or antibody fragment.
The first and second assays, used to select the conditionally active antibody or antibody fragment, are performed in assay solutions that may include a protein found in blood, such as albumin. The assays can also be performed in the absence of serum. The assay media used in the assays may include species with a molecular weight of less than 900 a.m.u. and a pKa up to 3 units away from the pH of 7.4 or between the pH of 6.0 and 7.4. Examples of species that can be included in the assay media are histidine, histamine, hydrogenated adenosine diphosphate, hydrogenated adenosine triphosphate, citrate, acetate, lactate, hydrogen sulfide, bisulfide, ammonium, bicarbonate, dihydrogen phosphate, and any combination thereof.
The method also allows for the selection of the conditionally active antibody or antibody fragment based on properties such as affinity, expression level, and humanization. The expression step can be performed using phage display or a eukaryotic cell production host, and if the latter is used, the selected conditionally active antibody or antibody fragment can be expressed in the same eukaryotic cell production host.
The acidic amino acid residues that can be introduced into the DNA are Aspartic acid, Glutamic acid, or a combination of both.
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GlobalData Patent Analytics tracks bibliographic data, legal events data, point in time patent ownerships, and backward and forward citations from global patenting offices. Textual analysis and official patent classifications are used to group patents into key thematic areas and link them to specific companies
