Point mutations are a type of gene mutation. point mutations include all individual base changes, i.e. deletions, insertions and substitutions. The effects of point mutations on the structure and function of amino acids and their constituent proteins are classified into three types, namely synonymous mutations, nonsense mutations and missense mutations. CD Formulation can mutate one or more sites of a protein molecule with a view to improving protein stability, increasing affinity or activity, reducing immunogenicity, etc. It is a key direction in protein drug design.
When the DNA double strand of a cell's genome is broken due to external factors, a DNA repair mechanism exists in the cell that is triggered to repair the DNA double strand at the broken site, which can be divided into homologous recombination repair and non-homologous recombination repair depending on the repair method. Homologous recombination means that DNA is repaired perfectly using a homologous DNA sequence as a template in the repair process, while non-homologous recombination repair means that the relevant proteins within the cell directly reattach the broken DNA ends, while new bases may be introduced randomly. The construction of point mutant cell lines is divided into three parts: (1) targeted DNA cleavage; (2) homologous recombination repair of DNA; and (3) cell screening.
CD Formulation can provide customers with mutant constructs to improve the stability, affinity, etc. of protein drugs. In addition, we can also provide the construction of anti-protease mutants and provide related testing services.
The main methods to extend the half-life of drugs by using constructs of mutants are as follows:
First, enhancing the degree of drug glycosylation as a way to increase the side chains on the drug surface, enhance protein smoothness and prevent rapid degradation of the drug by proteases;
Secondly, to increase the molecular weight of the drug to avoid glomerular filtration and prolong the action time of the free type drug.
Protease hydrolysis of biological drugs is an important barrier that needs to be broken in the development of drug delivery technology. Because proteins are readily broken down by proteases in endogenous serum or in tissues, and the degradation of proteins by proteases can occur at various stages of absorption, distribution, and excretion, this directly affects parameters such as the in vivo half-life, bioavailability, and plasma clearance of the drug. The introduction of one or more anti-protein hydrolysis mutants to biologically active molecules is the focus of research in drug design and development. In vivo stabilization is also achieved by the introduction of artificial amino acid analogues. These variants undergo small conformational changes leading to enhanced anti-protein hydrolysis.
Improving the stability of protein drugs against protease degradation by replacing arbitrary peptide bonds in the primary structure of proteins and changing the main structure of peptides by peptide backbone modification based on mimicking the biological activity of peptides are the goals of mimetic peptide design. To improve the ability of ribonuclease A to resist hydrolysis by proteinase K and Bacillus subtilis proteases, the conformation, biological activity and resistance to protease hydrolysis of the mutant enzymes were examined by targeted mutation, and later.
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