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CN-121851161-B - Alpha-syn specific antibody and application thereof in Parkinson auxiliary diagnosis kit

CN121851161BCN 121851161 BCN121851161 BCN 121851161BCN-121851161-B

Abstract

The application discloses an alpha-syn specific antibody and application thereof in an auxiliary diagnosis kit for parkinsonism, belongs to the technical field of immunoassay, and provides an antibody or antigen binding fragment thereof specifically binding to alpha-syn and a kit for detecting alpha-syn. The antibody of the application has high affinity, high sensitivity and obviously shortened production period, and is more suitable for being used as a core raw material in the field of in vitro diagnostic reagents. The kit disclosed by the application can be used for auxiliary diagnosis of diseases related to alpha-syn, such as Parkinson's disease.

Inventors

  • TANG BO
  • FENG SU
  • ZHENG DIWEN
  • WANG ZIMING
  • LI SHUANGFA

Assignees

  • 南京诺唯赞医疗科技有限公司
  • 湖南诺唯赞医疗科技有限公司

Dates

Publication Date
20260508
Application Date
20260316

Claims (10)

  1. 1. An antibody or antigen binding fragment thereof specifically binding to alpha-syn is characterized in that the amino acid sequence of heavy chain CDRH1-CDRH3 is shown as SEQ ID NO. 1-SEQ ID NO. 3, the amino acid sequence of light chain CDRL1 is shown as SEQ ID NO. 4, the amino acid sequence of light chain CDRL2 is EA, and the amino acid sequence of light chain CDRL3 is shown as SEQ ID NO. 5.
  2. 2. The antibody or antigen-binding fragment thereof according to claim 1, wherein it comprises a heavy chain variable region VH as shown in SEQ ID NO. 6 and a light chain variable region VL as shown in SEQ ID NO. 7.
  3. 3. A kit for detecting alpha-syn, which comprises magnetic beads coated by a first antibody and a second antibody marked by a chemiluminescent agent, wherein the second antibody in an antibody pair consisting of the first antibody and the second antibody is the antibody or antigen binding fragment thereof as claimed in claim 1.
  4. 4. The kit according to claim 3, wherein the amino acid sequence of the heavy chain CDRH1-CDRH3 of the first antibody is shown in SEQ ID NO. 17-SEQ ID NO. 19, the amino acid sequence of the light chain CDRL1 is shown in SEQ ID NO. 20, the amino acid sequence of the light chain CDRL2 is DA, and the amino acid sequence of the light chain CDRL3 is shown in SEQ ID NO. 21.
  5. 5. The kit according to claim 4, wherein the first antibody comprises a heavy chain variable region VH as shown in SEQ ID NO. 15 and a light chain variable region VL as shown in SEQ ID NO. 16.
  6. 6. The kit according to claim 3, wherein the chemiluminescent agent is at least one selected from the group consisting of acridinium esters, alkaline phosphatase, and horseradish peroxidase.
  7. 7. Use of the antibody or antigen binding fragment thereof of claim 1 in the preparation of a kit for detecting an α -syn protein.
  8. 8. Use of the antibody or antigen-binding fragment thereof of claim 1 in the preparation of a kit for aiding in the diagnosis of parkinson's disease.
  9. 9. A biomaterial, characterized by being selected from any one of the following: a. a polynucleotide encoding the antibody or antigen binding fragment thereof of claim 1; b. a vector comprising the polynucleotide of a; c. A host cell comprising b said vector or a polynucleotide of a integrated in the genome.
  10. 10. A kit for detecting α -syn, comprising the antibody or antigen-binding fragment thereof of claim 1.

Description

Alpha-syn specific antibody and application thereof in Parkinson auxiliary diagnosis kit Technical Field The application belongs to the technical field of immunoassay, and relates to an alpha-syn specific antibody and application thereof in an auxiliary diagnosis kit for parkinsonism. Background Parkinson's Disease (PD) is an age-related, progressive and irreversible neurodegenerative Disease, frequently occurring in the middle-aged and elderly. Clinically, it is manifested by symptoms of core movement such as bradykinesia, resting tremor, myotonia and dysposture balance, etc. in the absence of conscious disturbance, and various non-motor symptoms such as hyposmia, sleep disturbance, autonomic nerve dysfunction, cognitive dysfunction and depression anxiety, etc. Hoehn-Yahr stage is usually used to divide into 1-5 stages according to the degree of impaired exercise function and life self-care ability. The symptoms of patients in stage 1-2 are mostly limited to one side or two sides of the body, but the balance function is kept, the daily life can be self-care, the patients develop to stage 3, obvious instability and balance disorder of postures, the daily life begins to need partial assistance, the patients in stage 4-5 lose independent walking ability, need wheelchairs or lie on bed, and serious exercise complications and non-exercise symptoms appear. It was found that the main pathology of PD is manifested by progressive loss of the dopaminergic neurons of the substantia nigra compacta, and the appearance of lewis bodies (Lewy bodies) within the survivin neurons, which are predominantly responsible for abnormal aggregation of α -synuclein (α -synuclein), ultimately leading to dysfunction of the basal ganglia motor regulation loop. Alpha-syn is a normal protein encoded by the SNCA gene, present in all neurons, and its mutations, abnormal post-translational modifications or changes in the intracellular and extracellular environment, resulting in misfolding from a naturally disordered state, forming soluble oligomers. In physiological conditions, α -syn exists in neurons in a naturally disordered form, but in α -synucleinopathies such as PD, misfolding and dysfunction due to mutation, modification or environmental changes can lead to substantial reduction or loss of functional α -syn, and its normal physiological role is destroyed, triggering a series of downstream events including the formation of toxic oligomers, mitochondrial damage, inflammatory activation, and eventually aggregation into lewy bodies. Thus, the neurodegenerative process of PD results not only from the acquired toxicity of abnormal protein aggregates, but also from the progressive loss of normal α -syn function, which directly leads to loss of dopaminergic neurons and disturbance of basal ganglia, highlighting the fundamental role of α -syn itself in PD pathology. Thus, detecting α -syn levels may provide an important basis for clinical practice. Disclosure of Invention The application provides a specific antibody against alpha-syn, namely provides an alpha-syn specific antibody Anti-alpha-syn-rRmab-1. The monoclonal antibody prepared by adopting the single B cell technology has higher sensitivity and obviously shortened production period, and the kit prepared by utilizing the Anti-alpha-syn-rRmab-1 has high sensitivity and low detection limit, so that a tool is provided for auxiliary diagnosis of diseases related to alpha-syn, and the application of the antibody or the kit in auxiliary diagnosis of Parkinson is further provided. In one aspect, the application provides an antibody or antigen-binding fragment thereof that specifically binds to α -syn. In some embodiments, the antibody or antigen binding fragment comprises at least one, two, three, four, five, or six CDRs selected from (a) a heavy chain variable region CDR-H1 comprising, e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO:1, (b) a heavy chain variable region CDR-H2 comprising, e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO:2, (c) a heavy chain variable region CDR-H3 comprising, e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO:3, (d) a light chain variable region CDR-H3 comprising, e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO:4, and (e.g., 95%, 96%, 98% or 95% sequence identity to the amino acid sequence of 95%, 95% or 100% sequence of the amino acid sequence of SEQ ID NO: 3. In some embodiments, the antibody or antigen binding fragment comprises at least one, at least two, or all three VH CDR sequences selected from (a) comprising a heavy chain variable region CDR-H1 as having 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO