8 results about "Yeast display" patented technology

The invention discloses a nanometer antibody of mycobacterium tuberculosis early secretory protein ESAT-6 and application thereof, relates to the technical field of antibody recombination and genetic engineering antibodies, and is characterized by providing two nanometer antibodies AEN1 and AEN5 which are derived from a total synthesis alpaca nanometer antibody yeast display library and are combined with ESAT-6 in high affinity, DNA sequences for coding the AEN1 and the AEN5, a carrier containing the DNA, a carrier containing the carrier containing the DNA, and a carrier containing the carrier containing the carrier containing the DNA. The invention relates to a vector, a host cell containing the vector, and a method for preparing AEN1 and AEN5 by utilizing genetic engineering. And the method for quantitatively detecting the ESAT-6 by using the double-antibody sandwich ELISA is established. Detection on clinical specimens of tuberculosis shows that the sensitivity of the diagnosis method is 94.9%, and the specificity is 100%. The antibody is used for preparing gold-labeled rapid test paper, and the ESAT-6 can be rapidly detected from serum of a mycobacterium tuberculosis infected patient.

Disclosed are a nanobody targeting fibroblast activation protein-α, and the use thereof. An amino acid sequence of the nanobody comprises antigenic determinant complementary regions and framework regions with special structures. Firstly, alpacas are immunized with a recombinant FAPα antigen protein constructed to obtain a cell construct of alpaca PBMCs; secondly, an antibody targeting FAPα is screened by means of yeast display technology; and finally, an antibody having high sensitivity and specificity is obtained by means of functional detection and sequencing result analysis. The provided nanobody has the ability to specifically recognize and bind to FAPα, and has the characteristics of small molecular weight, low immunogenicity, better stability, a significant inhibitory effect on FAPα-expressing cells or tissues, etc., thus providing a potential therapeutic strategy for a disease involving FAPα expression. When the nanobody is used for developing or screening a diagnostic or therapeutic drug for FAPα-expressing cells or tissues, the obtained drug has excellent specificity and affinity, and has a significant inhibitory effect on FAPα-expressing cells or tissues.

The invention discloses a high-affinity HER2 targeting polycyclic peptide molecule and a preparation method and application thereof.The preparation method comprises the steps that affinity optimization is conducted on initial HER2 targeting polycyclic peptides obtained through phage display library screening by combining a yeast display technology with an error-prone PCR directed evolution system, and a series of high-affinity disulfide bond-rich polycyclic peptides (DDMP) are obtained; the polycyclic peptide is coupled with a conjugate (DOTA or FITC) and a linker, so that a monovalent or divalent molecular probe can be constructed; the molecular probe has high specificity, high stability and good biocompatibility, can be used for accurate imaging and quantitative evaluation of HER2 positive cells / tumors and diagnosis and targeted therapy of HER2 signal path related diseases through radionuclide labeling or fluorescence labeling, and provides a novel tool and a transformation basis for early diagnosis and accurate treatment of HER2 positive solid tumors.

The application discloses a method for modifying pH sensitivity of an antibody based on yeast display. A degenerate codon VAM (coding H, E, D, K, N, Q) is used to construct a yeast display library for at least two region combinations in LCDR1, HCDR1 and HCDR3, and 1-2 mutations are introduced into each CDR region, taking the antibody to be modified as a template. The library is transformed into Saccharomyces cerevisiae EBY100 to induce expression, and fast dissociation clones are enriched through four rounds of flow sorting (including pH 7.4 binding, pH 6.0 non-binding and'saturated binding-acid dissociation-neutral re-binding' screening). The obtained antibody has a KD ratio at pH 5.8 to pH 7.4 of up to 674.46, an expression amount of up to 263.3 mg / L, excellent physicochemical properties, and can enhance the curative effect of antibody conjugated drugs and improve the therapeutic index.

The polypeptide design method and sequence of the present application combine mask language modeling and yeast surface display, comprising the following steps: cleaning a protein sequence database, selecting protein sequences meeting the requirements as a training set of a language model, and performing mask language modeling on the protein sequences contained in the training set; designing a set of downstream tasks on the basis of a pre-trained model, and fine-tuning the downstream tasks; randomly masking residues of a selected reference sequence, and predicting the masked residues; performing virtual screening on polypeptide candidates generated by the model; and determining the protein expression level and affinity of the screened polypeptides through yeast display technology. The present application can generate a large number of polypeptide candidates that may have specific properties or functions by transferring natural language processing technology to the polypeptide generation field, and combines artificial intelligence generation, virtual screening and wet experimental characterization to build a relatively complete "dry-wet combination" design process.

It relates to a novel covalent protein, its pharmaceutical composition and use thereof in diagnosing, treating, or preventing diseases such as cancer and autoimmune disorders, which has the advantageous properties including high stability and potency, cost-effective production, enhanced tissue penetration, and the capacity for rapid and permanent target inhibition. It also relates to a screening method for developing covalent protein. It disclosure further details a high-throughput yeast display-based screening method for the development of such covalent proteins. It provides novel compounds as crosslinker for the covalent protein.

Via yeast display technology, albumin binding variants were selected, and variants displayed improved binding to MSA or both HSA & MSA. All variants were subcloned into the pHEN-expression plasmid and expressed in E coli WK6 cells. Purified nanobodies were then analyzed for their capacity to bind immobilized MSA. Biotinylated nanobodies were then tested in vivo for their circulatory half-life. They then validated the effect of their nanobodies anti albumin linked with another nanobodies such as single-domain antibody anti-D′D3 as described in WO2017129630 and WO2018091621.Mice having reduced levels of VWF (Von Willebrand disease type-1) received a single dose of the bispecific single-domain antibody. VWF and FVIII levels were followed over 14 days. These data show that there is a statistically significant increase in VWF and FVIII levels, which is maintained for 10 days.The present invention relates to an isolated single-domain antibody (sdAb) directed against albumin and its uses in different diseases such as bleeding disorders.

The present invention relates to a nanoantibody targeting phosphatidylinositol proteoglycan 3 and the use thereof. The amino acid sequence of the nanoantibody comprises an antigenic complementarity determining region and a framework region of a special structure. First, immunizing a llama by means of constructing GPC3 antigens so as to obtain llama PBMC cell constructs; then, screening out GPC3-targeting antibodies by means of a yeast display technique; and finally, obtaining an antibody having high sensitivity and specificity by means of functional detection and sequencing result analysis. The provided nanoantibody has specific recognition and binding ability for GPC3, has a small molecular weight, low immunogenicity, high stability, and a significant tumor inhibition effect, and provides a potential strategy for treating GPC3-expressing tumor diseases. The present invention is used for developing or screening for a drug for the diagnosis or treatment of GPC3-expressing cells or tumors, and the obtained drug has good specificity and affinity and a significant tumor inhibition effect.