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Monoclonal antibody against echo30, preparation method and application thereof

A technology of monoclonal antibody and antigen, applied in the field of biomedicine, can solve the problems of non-specific antibody reports, less Echo30 virus, etc.

Active Publication Date: 2020-10-27
JIANGSU PROVINCIAL CENT FOR DISEASE CONTROL & PREVENTION PUBLIC HEALTH RES INST OF JIANGSU PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are relatively few studies on Echo30 virus, and there are no reports of specific antibodies

Method used

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  • Monoclonal antibody against echo30, preparation method and application thereof
  • Monoclonal antibody against echo30, preparation method and application thereof
  • Monoclonal antibody against echo30, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Embodiment 1 monoclonal antibody preparation

[0089] 1. Immunogen (antigen): Echo30.

[0090] Each mouse was immunized 5 times, each time with 100 μl of antigen, and a total of 3-3.5ml of virus was needed.

[0091] 2. Animal immunity

[0092] Two adjuvants were used for immunization, namely modified Freund's adjuvant and aqueous adjuvant, two groups were immunized with virus, and each group of 3 mice was used as one adjuvant, and a total of 6 mice were immunized with virus. All immunizations were performed in SPF animal rooms.

[0093] Select 6 Balb / c mice (female mice) aged 5-8 weeks, mix the immunogen and adjuvant and then emulsify them separately for immunization. Freund's complete adjuvant is used for the first injection, and Freund's complete adjuvant is used for the next 4 booster injections. Incomplete adjuvant, mixed well with the same volume of antigen before injection.

[0094] The immunization method is multi-point injection on the back, the main injecti...

Embodiment 2

[0121] Example 2 Antibody function

[0122] 1. Antigen-antibody affinity determination

[0123] 1.1 Surface plasmon resonance (SPR)

[0124] 1.1.1 Steps:

[0125] Cut and purify 4B10-IgG (full-length antibody) and 6C5-IgG (full-length antibody) using Pierce Fab Preparation Kit (Thermo) to obtain 4B10-Fab (Fab fragment of antibody) and 6C5-Fab (Fab fragment of antibody) .

[0126] SPR experiments were performed using BIAcore T100 (Biacore, GE Healthcare), and the buffer was PBS containing 0.05% Tween-20. Purified E30 whole virus particles were immobilized on the surface of a CM5 sensor chip using the NHS / EDC method until the RU value reached 740. Gradient concentrations of IgG or Fab were then flowed through the chip at a rate of 20 μl / min, and the chip was regenerated using 10 mM glycine-HCl (pH 1.7) after each injection cycle. Binding affinities were obtained by global fitting of curves using the software BIAevaluation (version 4.1).

[0127] 1.1.2 Results

[0128] The...

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Abstract

The invention discloses a monoclonal antibody for Echo30 and a preparation method and application of the monoclonal antibody. The invention further discloses an antigen binding fragment of an anti-Echo30 antibody. The invention further relates to a composition, a detection product and a diagnosis product with the antibody. The invention further relates to separated nucleic acid for encoding an antibody sequence, a carrier, a host cell and application of the antibody in diagnosis or treatment.

Description

technical field [0001] The application belongs to the field of biomedicine, and specifically relates to a monoclonal antibody against Echo30, a preparation method and an application thereof. Background technique [0002] Human intestinal cytopathogenic orphan virus (Enteric Cytopathogenic human Orphan Virus, ECHO), referred to as Echo virus, is a type of single-stranded positive-sense RNA enterovirus, with 34 serotypes. Its pathogenicity was not clear at first, but it was later proved that echovirus was related to various diseases such as aseptic meningitis, infant diarrhea, hand, foot and mouth disease. A large number of studies and data analysis have finally shown that the disease spreads all over the world and causes widespread spread or epidemic among the population. Its infection can cause various human diseases such as aseptic meningitis, rash, gastrointestinal diseases, hepatitis and pneumonia, etc. Among them, aseptic meningitis is the most common and can be transmi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10C12N15/41C12N15/85G01N33/577G01N33/569A61K39/42A61P31/14
CPCA61K2039/505A61P31/14C07K16/1009C07K2317/565C12N15/85G01N33/56983G01N33/577G01N2333/085
Inventor 张黎王康郑滨洋朱玲苏璇张倩陶焱炀崔仑标潘红星葛以跃吴涛王祥喜朱凤才
Owner JIANGSU PROVINCIAL CENT FOR DISEASE CONTROL & PREVENTION PUBLIC HEALTH RES INST OF JIANGSU PROVINCE