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CN-115197294-B - Polypeptide, polypeptide composition, kit and related application

CN115197294BCN 115197294 BCN115197294 BCN 115197294BCN-115197294-B

Abstract

The invention provides a polypeptide, a polypeptide composition, a kit and related applications. The polypeptide is shown as SEQ ID NO. 1-SEQ ID NO. 85, and can specifically bind to a target antibody, such as a human SNRPC/U1C antibody, a human PCNA antibody, a human CENPB antibody, a human NUP210/gp210 antibody, a human FTCD/58K Golgi protein antibody and a GAD antibody. The above-mentioned multiple polypeptides or polypeptide sets specifically binding to the target antibody may cover peptide segments including linear epitopes and nonlinear epitopes, so that it is convenient to reasonably select the number of the used polypeptides according to the number of the target antibodies actually to be detected, so as to improve the detection sensitivity and specificity of each polypeptide, improve the detection accuracy, and improve the flux and efficiency of the detected antibodies.

Inventors

  • XIE CHUNLAN
  • GUO BAOSEN
  • LIU YING
  • ZHENG HANCHENG
  • LI DANNI
  • ZHANG JUNJIE
  • Gong Zhuolin

Assignees

  • 珠海碳云智能科技有限公司

Dates

Publication Date
20260508
Application Date
20210408

Claims (13)

  1. 1. A polypeptide composition comprising one or more polypeptides capable of specifically binding one or more antibodies of interest selected from any one of human SNRPC/U1C antibody, human SNRPC/U1C antibody + human CENPB antibody, human SNRPC/U1C antibody + human CENPB antibody + human PCNA or human SNRPC/U1C antibody + human CENPB antibody + human PCNA antibody + human NUP 210/gp 210; When the target antibody is human SNRPC/U1C antibody, the polypeptide is selected from any one or more of SEQ ID NO. 1 to SEQ ID NO. 7 and SEQ ID NOs 9-10; When the target antibody is an antibody of human PCNA, the polypeptide is selected from any one or more of SEQ ID NO. 11 to SEQ ID NO. 15; When the target antibody is human CENPB, the polypeptide is selected from any one or more of SEQ ID NO. 16 to SEQ ID NO. 26; When the target antibody is an antibody of human NUP 210/gp 210, the polypeptide is selected from any one or more of SEQ ID NO. 27 to SEQ ID NO. 35.
  2. 2. A polypeptide product comprising one or more polypeptides capable of specifically binding one or more antibodies of interest selected from any one of human SNRPC/U1C antibody, human SNRPC/U1C antibody + human CENPB antibody, human SNRPC/U1C antibody + human CENPB antibody + human PCNA or human SNRPC/U1C antibody + human CENPB antibody + human PCNA antibody + human NUP 210/gp 210 antibody; When the target antibody is human SNRPC/U1C antibody, the polypeptide is selected from any one or more of SEQ ID NO. 1 to SEQ ID NO. 7 and SEQ ID NOs 9-10; When the target antibody is an antibody of human PCNA, the polypeptide is selected from any one or more of SEQ ID NO. 11 to SEQ ID NO. 15; When the target antibody is human CENPB, the polypeptide is selected from any one or more of SEQ ID NO. 16 to SEQ ID NO. 26; When the target antibody is an antibody of human NUP 210/gp 210, the polypeptide is selected from any one or more of SEQ ID NO. 27 to SEQ ID NO. 35.
  3. 3. The polypeptide product of claim 2, wherein the polypeptide product further comprises a polypeptide stabilizer.
  4. 4. The polypeptide product of claim 3, wherein the polypeptide stabilizer comprises 150-180 mm NaCl, 100-140 mm polylysine hydrochloride, and water; the polypeptide product is a polypeptide chip, and the polypeptides on the polypeptide chip consist of the polypeptides.
  5. 5. An antibody detection reagent comprising one or more polypeptides, wherein one or more of said polypeptides is capable of specifically binding one or more antibodies of interest selected from any one of human SNRPC/U1C antibody, human SNRPC/U1C antibody+human CENPB antibody, human SNRPC/U1C antibody+human CENPB antibody+human PCNA antibody or human SNRPC/U1C antibody+human CENPB antibody+human PCNA antibody+human NUP 210/gp 210 antibody; When the target antibody is human SNRPC/U1C antibody, the polypeptide is selected from any one or more of SEQ ID NO. 1 to SEQ ID NO. 7 and SEQ ID NOs 9-10; When the target antibody is an antibody of human PCNA, the polypeptide is selected from any one or more of SEQ ID NO. 11 to SEQ ID NO. 15; When the target antibody is human CENPB, the polypeptide is selected from any one or more of SEQ ID NO. 16 to SEQ ID NO. 26; When the target antibody is an antibody of human NUP 210/gp 210, the polypeptide is selected from any one or more of SEQ ID NO. 27 to SEQ ID NO. 35.
  6. 6. An antibody detection kit, characterized in that the kit contains a plurality of polypeptides, wherein the polypeptides can specifically bind one or more target antibodies, and one or more target antibodies are selected from any one of human SNRPC/U1C antibody, human SNRPC/U1C antibody+human CENPB antibody, human SNRPC/U1C antibody+human CENPB antibody+human PCNA antibody or human SNRPC/U1C antibody+human CENPB antibody+human PCNA antibody+human NUP 210/gp 210 antibody; when the target antibody is human SNRPC/U1C antibody, the polypeptide is selected from any one or more of SEQ ID NO.1 to SEQ ID NO. 7 and SEQ ID NO. 9-10; When the target antibody is an antibody of human PCNA, the polypeptide is selected from any one or more of SEQ ID NO. 11 to SEQ ID NO. 15; When the target antibody is human CENPB, the polypeptide is selected from any one or more of SEQ ID NO. 16 to SEQ ID NO. 26; When the target antibody is an antibody of human NUP 210/gp 210, the polypeptide is selected from any one or more of SEQ ID NO. 27 to SEQ ID NO. 35.
  7. 7. The kit of claim 6, wherein the kit comprises a detection chip, the plurality of polypeptides is disposed on the detection chip, and the polypeptides on the detection chip consist of the plurality of polypeptides.
  8. 8. The use of the polypeptide composition according to claim 1 for preparing a kit for detecting antibodies, wherein the antibodies are selected from any one or more of human SNRPC/U1C antibodies, human CENPB antibodies, human PCNA antibodies or human NUP 210/gp 210 antibodies.
  9. 9. The use of the polypeptide composition of claim 1 for the preparation of a product for detecting antibodies, wherein the antibodies are selected from any one or more of the group consisting of antibodies to human SNRPC/U1C, human CENPB, human PCNA or human NUP 210/gp 210.
  10. 10. The use according to claim 9, wherein the detection antibody product further comprises a sample diluent which is a PBST buffer containing D-mannitol, wherein the mass volume content of D-mannitol in the PBST buffer is 0.5% -1%.
  11. 11. The use according to claim 10, wherein the detection antibody product further comprises a blocking solution comprising 130-137 mM sodium chloride, 2.5-2.7 mM potassium chloride, 3.8-4.3 mM disodium hydrogen phosphate, 1.2-1.4 mM potassium dihydrogen phosphate, 0.05-1% Tween-20 v/v, 0.05-0.1% Proclin950 v/v, 0.5-1% D-mannitol w/v and 0.1-1% casein w/v, the blocking solution having a pH of 7.2-7.6.
  12. 12. The use according to claim 11, wherein the pH of the confining liquid is 7.38-7.42.
  13. 13. The use according to claim 12, wherein the blocking solution is 137 mM sodium chloride, 2.7 mM potassium chloride, 4.3 mM disodium hydrogen phosphate, 1.4 mM potassium dihydrogen phosphate, 1% Tween-20 v/v, 0.1% Proclin950 v/v, 1% D-mannitol w/v and 0.1% casein w/v, and the pH of the blocking solution is 7.4.

Description

Polypeptide, polypeptide composition, kit and related application Technical Field The invention relates to the field of antibody detection, in particular to a polypeptide, a polypeptide composition, a kit and related applications. Background In immune response, cellular immunity and humoral immunity are two closely related and mutually regulated physiological processes, and in clinical examination work, detection by specific antibodies in humoral immune response is most widely used. The specific antibody detection has important clinical significance, can assist clinical diagnosis, for example, the detection of the specific antibody in serum or blood plasma can be used as an auxiliary reference for clinical diagnosis of autoimmune diseases, infectious diseases, allergic diseases, tumors and other diseases, and is an index for observing curative effects and prognosis in certain diseases. In addition, the observation of vaccination effect is also of special and important significance in the investigation of epidemiology of infectious diseases. At present, a plurality of antibody detection methods are adopted, and besides the traditional precipitation reaction, agglutination test and complement fixation test, the marked immunoassay such as enzyme-linked immunoassay, radioimmunoassay, fluorescent immunoassay, luminescent immunoassay and the like are already the main immunoassay technologies, and immunoblotting and rapid spot immunobinding test are also widely used. However, these conventional methods generally detect only one antibody index in one experiment, and have low detection sensitivity and large sample usage. Recently, detection of antibodies by the polypeptide chip method has also been reported. However, this method has some problems in clinical use in detection of specific antibodies against common diseases, such as: 1) The polypeptide chip designed based on the antigen sequence information can detect the linear epitope more clearly, and correspondingly, the epitope corresponding peptide segment can be used for antibody detection. However, this method cannot meet the detection requirements for ubiquitous nonlinear epitopes. While there are currently epitopes that constrain polypeptides by one or more disulfide bonds, which mimic discontinuities and are conformation dependent, the possibility of detection of only a small number of non-linear epitopes is still increased. 2) The number of peptide fragments of the random sequence polypeptide chip is large, the target antibody conformational epitope core recognition site can be only a few amino acids, and the antibody can be combined when 4-5 amino acid residues in the peptide fragments are perfectly matched, so that the signal peptide fragments after the random sequence chip is incubated with the antibody can contain 1) an antigen epitope comprising a linear epitope and/or a conformational epitope, 2) a sequence containing the core recognition amino acid of the linear epitope and/or the conformational epitope, 3) a non-specific combined peptide fragment and the like, but a non-specific interference signal can not be removed through a sequence comparison equivalent method. 3) The antibodies in the organism are polyclonal antibodies, the antigen epitope is not unique, and the antibody detection is easy to miss by using the peptide fragment to simulate a single epitope. 4) The clinical antibody detection mainly uses serum/plasma samples, the components are complex, and the specific peptide fragments obtained by analysis after incubation of pure antibodies on a polypeptide chip can be interfered by other proteins in the serum/plasma. 5) The clinical antibodies are of various kinds (for example, there are various kinds of antibodies to be detected for infectious diseases, allergic diseases, tumors, etc., and autoimmune diseases are exemplified by autoimmune diseases, and one autoimmune disease can produce a plurality of autoimmune antibodies, and the same autoimmune antibody can exist in a plurality of autoimmune diseases), and the detection frequency is also high. Thus, there remains a need to provide new specific antibody detection products, which address the above-mentioned clinical detection needs. Disclosure of Invention The invention mainly aims at providing a polypeptide, a polypeptide composition, a kit and related applications, so as to provide an antibody detection product with higher specificity. In order to achieve the above object, according to one aspect of the present invention, there is provided a polypeptide capable of specifically binding to a target antibody selected from any one or more of human SNRPC/U1C antibody, human PCNA antibody, human CENPB antibody, human NUP210/gp210 antibody, human FTCD/58K Golgi protein antibody and GAD antibody, wherein the polypeptide is selected from any one or more of SEQ ID NO. 1 to SEQ ID NO. 10 when the target antibody is human SNRPC/U1C antibody, the polypeptide is selected from any one or more of SEQ ID N