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CN-121991217-A - Antibody, antibody pair, reagent and method for detecting glial fibrillary acidic protein

CN121991217ACN 121991217 ACN121991217 ACN 121991217ACN-121991217-A

Abstract

The invention discloses an antibody, an antibody pair, a reagent and a method for detecting glial fibrillary acidic protein, and relates to the field of immunodiagnosis. The antibody pair for detecting the glial fibrillary acidic protein disclosed by the invention comprises the first antibody and the second antibody, and the reagent and the detection method based on the antibody and the antibody pair can accurately detect the presence of the glial fibrillary acidic protein.

Inventors

  • GUO WEIYUN
  • LI ZHEN
  • CHEN XIAOYUN

Assignees

  • 东莞市朋志生物科技有限公司

Dates

Publication Date
20260508
Application Date
20241101

Claims (12)

  1. 1. An antibody pair for detecting glial fibrillary acidic protein, characterized in that, The antibody pair comprises a first antibody and a second antibody, wherein the first antibody comprises three heavy chain complementarity determining regions (HCDR 1, HCDR2 and HCDR 3) in the heavy chain variable region shown in SEQ ID NO. 9 and three light chain complementarity determining regions (LCDR 1, LCDR2 and LCDR 3) in the light chain variable region shown in SEQ ID NO. 11; The second antibody comprises three heavy chain complementarity determining regions HCDR1, HCDR2 and HCDR3 in the heavy chain variable region shown in SEQ ID NO. 19 and three light chain complementarity determining regions LCDR1, LCDR2 and LCDR3 in the light chain variable region shown in SEQ ID NO. 21; Optionally, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, LCDR3 is defined by any one or a combination of systems Kabat, chothia, IMGT, abM or contacts.
  2. 2. An antibody pair for detecting glial acidic protein, characterized in that the antibody pair comprises a first antibody and a second antibody, wherein the first antibody has HCDR1 shown as SEQ ID NO. 1 (FYIIH), HCDR2 shown as SEQ ID NO. 2 (FINPSNTYTNYNHKFKD), HCDR3 shown as SEQ ID NO. 3 (AQFAY), LCDR1 shown as SEQ ID NO. 4 (KSSQSLLHSDGKTYLN), LCDR2 shown as SEQ ID NO. 5 (LVSKLDS), LCDR3 shown as SEQ ID NO. 6 (WQGTHFPWT); The second antibody has HCDR1 shown in SEQ ID NO. 13 (DYWMH), HCDR2 shown in SEQ ID NO. 14 (AIDTSTSYTTNNQKFKG), HCDR3 shown in SEQ ID NO. 15 (SAYYFDY), LCDR1 shown in SEQ ID NO. 16 (KSSQSLLNSANQKNYLA), LCDR2 shown in SEQ ID NO. 17 (FASTRES) and LCDR3 shown in SEQ ID NO. 18 (QQHYITPFT).
  3. 3. The antibody pair for detecting glial fibrin according to any one of claims 1 to 2, wherein the first and second antibodies further comprise framework regions indicated by HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3 and LFR 4; Optionally, the first antibody comprises HFR1, HFR2, HFR3, HFR4 with amino acid sequences shown in SEQ ID NO.23 to SEQ ID NO. 26, and LFR1, LFR2, LFR3, LFR4 with amino acid sequences shown in SEQ ID NO. 27 to SEQ ID NO. 30, or an amino acid sequence having at least 80% identity to each of the framework region sequences; The second antibody comprises HFR1, HFR2, HFR3 and HFR4 with amino acid sequences shown in SEQ ID NO. 31 to SEQ ID NO. 34 and LFR1, LFR2, LFR3 and LFR4 with amino acid sequences shown in SEQ ID NO. 35 to SEQ ID NO. 38 or amino acid sequences with at least 80% identity with the sequences of the framework regions.
  4. 4. An antibody for detecting glial fibrillary acidic protein, characterized in that the complementarity determining region of the antibody comprises any one of (a) - (b): (a) The amino acid sequences are shown as HCDR1, HCDR2 and HCDR3 shown in SEQ ID NO 1 to SEQ ID NO 3 in sequence, and the amino acid sequences are shown as LCDR1, LCDR2 and LCDR3 shown in SEQ ID NO 4 to SEQ ID NO 6 in sequence; (b) The amino acid sequences are shown as HCDR1, HCDR2 and HCDR3 shown in SEQ ID NO 13 to SEQ ID NO 15 in sequence, and the amino acid sequences are shown as LCDR1, LCDR2 and LCDR3 shown in SEQ ID NO 16 to SEQ ID NO 18 in sequence; Optionally, the antibody further comprises a framework region shown as HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR 4; optionally, the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 are selected from any one of (a ') to (b'): (a') HFR1, HFR2, HFR3, HFR4 having amino acid sequences shown in SEQ ID NO. 23 to SEQ ID NO. 26, and LFR1, LFR2, LFR3, LFR4 having amino acid sequences shown in SEQ ID NO. 27 to SEQ ID NO. 30, or an amino acid sequence having at least 80% identity to each of said framework region sequences; (b') HFR1, HFR2, HFR3, HFR4 having the amino acid sequences shown in SEQ ID NO. 31 to SEQ ID NO. 34, and LFR1, LFR2, LFR3, LFR4 having the amino acid sequences shown in SEQ ID NO. 35 to SEQ ID NO. 38, or an amino acid sequence having at least 80% identity to the sequence of each of the framework regions.
  5. 5. An antibody pair for detecting glial fibrillary acidic protein, characterized in that, The antibody pair comprises a first antibody and a second antibody, wherein the first antibody comprises a heavy chain variable region with an amino acid sequence shown as SEQ ID NO. 9 and a light chain variable region with an amino acid sequence shown as SEQ ID NO. 11; the second antibody comprises a heavy chain variable region with an amino acid sequence shown as SEQ ID NO. 19 and a light chain variable region with an amino acid sequence shown as SEQ ID NO. 21.
  6. 6. An antibody for detecting glial fibrillary acidic protein, which is characterized by comprising a heavy chain variable region and a light chain variable region, wherein the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 9 or 19, and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 11 or 21.
  7. 7. The antibody pair for detecting glial fibrillary acidic protein according to any one of claims 1 to 3, 5, or the antibody of claim 4 or 6, wherein the first antibody and the second antibody of the antibody pair or the antibody further comprises a constant region; Alternatively, the constant region is of a species source of bovine, equine, porcine, ovine, caprine, rat, mouse, canine, camel, feline, rabbit, donkey, deer, mink, chicken, duck, goose, or human; alternatively, the constant region is of mouse species origin; optionally, the constant region comprises a heavy chain constant region and/or a light chain constant region; optionally, the heavy chain constant region is selected from the group consisting of a heavy chain constant region of any one of IgG1, igG2, igG3, igG4, igA, igM, igE, igD, or a combination of constant regions, and/or the light chain constant region is selected from the group consisting of kappa-type or lambda-type light chain constant regions; Optionally, the heavy chain constant region comprises CH1 of IgG1, a hinge region of IgG1, CH2 of IgM, CH3 of IgM, and/or CH4 of IgM; Alternatively, the heavy chain constant region of the first, second or antibody is SEQ ID NO. 7 or a sequence having at least 80% identity thereto and the light chain constant region of the first, second or antibody is SEQ ID NO. 8 or a sequence having at least 80% identity thereto.
  8. 8. An antibody pair for detecting glial fibrillary acidic protein, characterized in that the antibody pair comprises a first antibody and a second antibody, The first antibody comprises a heavy chain with an amino acid sequence shown as SEQ ID NO. 10 and a light chain with an amino acid sequence shown as SEQ ID NO. 12, and the second antibody comprises a heavy chain with an amino acid sequence shown as SEQ ID NO. 20 and a light chain with an amino acid sequence shown as SEQ ID NO. 22.
  9. 9. An antibody for detecting glial fibrillary acidic protein, which is characterized by comprising a heavy chain and a light chain, wherein the amino acid sequence of the heavy chain is shown as SEQ ID NO. 10 or 20, and the amino acid sequence of the light chain is shown as SEQ ID NO. 12 or 22.
  10. 10. A reagent or kit for detecting a glial acidic protein, characterized in that the reagent or kit comprises the antibody pair for detecting a glial acidic protein of any one of claims 1 to 3, 5, 7, 8 or the antibody of claims 4, 6, 7, 9; optionally, the first antibody in the antibody pair is a coated antibody, and the second antibody is a labeled antibody; optionally, the second antibody in the antibody pair is a coated antibody, and the first antibody is a labeled antibody; Optionally, the antibody is a labeled antibody or a coated antibody; Optionally, the labeled antibody is labeled with a detectable label and/or binding partner; optionally, the detectable label is selected from the group consisting of metal ions, fluorescent dyes, enzymes, radioisotopes, chemiluminescent labels, electron dense labels, adamantane, and nanoparticle-based labels; optionally, the coated antibody is attached to a solid phase and/or a binding partner; Optionally, the solid phase is selected from microspheres, magnetic beads, latex particles, microfluidic chips, plates or membranes; Alternatively, the binding partner comprises a biotin/avidin or biotin derivative/avidin derivative protein.
  11. 11. A method for detecting glial fibrillary acidic protein, comprising: a) Contacting the antibody pair for detecting glial fibrillary acidic protein of any one of claims 1 to 3, 5, 7, 8, or the antibody of any one of claims 4, 6, 7, 9, or the reagent or kit of claim 11 with a sample to be detected under conditions sufficient for an antibody/antigen binding reaction to form an immune complex, and B) Detecting the presence of the immune complex, the presence of the complex being indicative of the presence of glial fibrillary acidic protein in the test sample.
  12. 12. Use of an antibody pair for detecting glial fibrillary acidic protein according to any one of claims 1 to 3, 5, 7, 8, or an antibody according to any one of claims 4, 6, 7, 9, or a reagent or kit according to claim 11 for detecting glial fibrillary acidic protein or for preparing a product for detecting glial fibrillary acidic protein.

Description

Antibody, antibody pair, reagent and method for detecting glial fibrillary acidic protein Technical Field The invention relates to the technical field of immunodiagnosis, in particular to an antibody, an antibody pair, a reagent and a method for detecting glial fibrillary acidic protein. Background Traumatic brain injury (Traumatic brain injury, TBI) refers to brain injury caused by external shock or vibration, resulting in degradation or damage of cognitive function or physical function. TBI accounts for 17% -23% of all wounds, the death rate of severe craniocerebral injury is more than 20%, the serious disability rate is more than 50%, and the death rate is the first. It was investigated that 22% of TBI patients under 15 years old were caused by foreign objects striking the head, and 79% of TBI patients over 65 years old were caused by accidental falls. The current method for diagnosing TBI mainly carries out preliminary injury judgment according to the Grassgo coma score (Glasgow Coma Scale, GCS) and the coma time, and clinical laboratory diagnosis mainly depends on cranium CT (computed tomography of the head), nuclear magnetic resonance imaging MRI (Magnetic Resonance Imaging) and the like. Although these methods play an important role in diagnosis, they also have some significant drawbacks such as long time consumption, insufficient sensitivity, high radiation exposure risk, and the like. In recent years, diagnosis of brain injury by biomarkers has become a research hotspot, wherein glial fibrillary acidic protein (Glial Fibrillary Acidic Protein, GFAP) has been of great interest as a potential biomarker due to its correlation with TBI severity. GFAP is an astrocyte-expressed cytoskeletal protein that is released into the blood after brain injury, and changes in its levels can be used to assess the extent of nerve injury. Thus, GFAP level changes are of great clinical value for early diagnosis and risk assessment of brain injury diseases. The immune detection method is widely applied to detection of the biomarker due to simple operation, high sensitivity and strong specificity. Common immunodetection methods include a colloidal gold method, a fluorescence method, a chemiluminescence method and the like, and the main limitations of the immunodetection methods are that antibody raw materials aiming at GFAP are lacked, in addition, paired antibodies of GFAP proteins are key for realizing immunodetection of the GFAP proteins besides the antibody raw materials, and antibody pairing with performance advantages is lacked in the market, which limits the accuracy and reliability of GFAP detection, and strong demands are made in the field for antibodies and antibody pairing with high sensitivity and accuracy. In addition, in the case of the optical fiber, Disclosure of Invention The application provides an antibody and an antibody pair, which provide an important raw material source for detecting the glial fibrillary acidic protein and have good detection performance. In order to achieve the above object, according to a first aspect of the present invention, there is provided an antibody pair for detecting glial fibrillary acidic protein, The antibody pair comprises a first antibody and a second antibody, wherein the first antibody comprises three heavy chain complementarity determining regions (HCDR 1, HCDR2 and HCDR 3) in the heavy chain variable region shown in SEQ ID NO. 9 and three light chain complementarity determining regions (LCDR 1, LCDR2 and LCDR 3) in the light chain variable region shown in SEQ ID NO. 11; the second antibody comprises three heavy chain complementarity determining regions HCDR1, HCDR2 and HCDR3 in the heavy chain variable region shown in SEQ ID NO. 19 and three light chain complementarity determining regions LCDR1, LCDR2 and LCDR3 in the light chain variable region shown in SEQ ID NO. 21. In order to achieve the above object, according to a second aspect of the present invention, there is provided an antibody pair for detecting glial fibrillary acidic protein, The antibody pair comprises a first antibody and a second antibody, wherein the first antibody has HCDR1 shown as SEQ ID NO. 1 (FYIIH), HCDR2 shown as SEQ ID NO. 2 (FINPSNTYTNYNHKFKD), HCDR3 shown as SEQ ID NO. 3 (AQFAY), LCDR1 shown as SEQ ID NO. 4 (KSSQSLLHSDGKTYLN), LCDR2 shown as SEQ ID NO. 5 (LVSKLDS) and LCDR3 shown as SEQ ID NO. 6 (WQGTHFPWT); The second antibody has HCDR1 shown in SEQ ID NO. 13 (DYWMH), HCDR2 shown in SEQ ID NO. 14 (AIDTSTSYTTNNQKFKG), HCDR3 shown in SEQ ID NO. 15 (SAYYFDY), LCDR1 shown in SEQ ID NO. 16 (KSSQSLLNSANQKNYLA), LCDR2 shown in SEQ ID NO. 17 (FASTRES) and LCDR3 shown in SEQ ID NO. 18 (QQHYITPFT). In order to achieve the above object, according to a third aspect of the present invention, there is provided an antibody for detecting glial fibrillary acidic protein, the complementarity determining region of the antibody comprising any one of (a) - (b): (a) The amino acid sequences are shown as HC