4results about How to "Strong binding specificity" patented technology

This invention discloses a method for preparing chlorantraniliprole nanobodies, belonging to the field of biorecognition material preparation for pesticide residue detection. The method involves screening chlorantraniliprole-specific nanobodies using bioinformatics analysis, cloning them into the pET-22b(+) expression vector, transforming them into *E. coli* BL21(DE3) competent cells, and achieving soluble expression of the nanobodies via IPTG induction. High-purity chlorantraniliprole nanobodies are then obtained through periplasmic purification combined with nickel column affinity chromatography. This invention eliminates the need for animal immunization experiments such as alpaca screening through bioinformatics, significantly shortening the nanobodies preparation cycle and reducing costs. The use of the pET-22b(+) prokaryotic expression system combined with an 18℃ low-temperature induction strategy achieves soluble expression of chlorantraniliprole nanobodies. After induction, the target protein band can be clearly detected by SDS-PAGE electrophoresis (see Figure 1), improving the correct folding ratio of the antibody.

This invention discloses a flow cytometry method for detecting fluorescent microspheres based on TSA signal amplification. The method includes: incubating fluorescently encoded microspheres coated with a capture antibody with a sample; adding a biotinylated detection antibody to form a sandwich complex; subsequently introducing two high-density signal amplification polymers sequentially: SA-PolyHRP400 as the first signal amplification polymer, and a second signal amplification polymer with a polysaccharide backbone and covalently coupled with multiple tyrosine and luciferin molecules. Under HRP catalysis, the tyrosine is activated and covalently deposited in the vicinity of the microspheres, achieving exponential amplification of the fluorescence signal. This method is simple to operate, more efficient than traditional methods, and more suitable for high-throughput, high-timeliness platforms like flow cytometry. It also exhibits high sensitivity, high specificity, and good repeatability for detecting low-abundance proteins such as IL-6 based on the sandwich method, and is compatible with non-magnetic / magnetic fluorescently encoded microspheres of various sizes. It is suitable for multi-indicator detection and can be widely applied to early clinical diagnosis and trace biomarker analysis.

This disclosure provides a PD-1 antibody and its application. The PD-1 antibody or antigen-binding moiety provided in this disclosure has a high affinity for human PD-1 protein and can bind at a rate of 0.9 x 10⁻⁶. ‑8 The Kd value of M binds to human PD-1. Furthermore, the binding specificity is strong, and it does not cross-react with human CTLA-4 positive cells (3T3 cells overexpressing human CTLA-4) or human PD-1 negative cells (such as Raji cells). Based on these characteristics, the PD-1 antibody or antigen-binding moiety provided in this disclosure can be used for the detection of human PD-1 protein, and can also be used alone or in combination with other methods in tumor immunotherapy. That is, it can be effectively applied in the preparation of drugs for treating tumors, infectious diseases, autoimmune diseases, and anti-immune rejection.

This invention belongs to the field of phosphate binders, specifically relating to a linear saturated aliphatic dicarboxylic acid-iron(III) complex, its preparation method, and its applications, particularly its use in preparing drugs for the treatment or prevention of hyperphosphatemia. In the dicarboxylic acid-iron(III) complex, the molar ratio of iron to dicarboxylic acid is 1:0.8~1.2, and the dicarboxylic acid is a linear saturated aliphatic dicarboxylic acid. The complex is prepared by reacting a ferric salt and a dicarboxylic acid salt in a solvent. In vitro phosphate binding experiments demonstrate that the dicarboxylic acid-iron(III) complex of this invention exhibits high phosphate binding capacity per unit mass of iron within the pH range of 3-7, and maintains a high phosphate binding capacity per unit mass of iron in vivo, significantly superior to commercially available sucrose ferric hydroxide chewable tablets. As an oral phosphate binder, the complex of this invention exhibits stronger phosphate binding capacity per unit mass of iron in vivo, higher iron utilization, lower total iron intake, and fewer adverse reactions.