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Neutralizing antibody against respiratory syncytial viruses and application thereof

A syncytial virus and anti-respiratory technology, applied in the fields of medicine and immunology, can solve problems such as severe toxicity and teratogenicity

Active Publication Date: 2019-07-16
ZHUHAI TRINOMAB BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, palivizumab is an expensive humanized monoclonal antibody that can only be used as a preventive treatment; ribavirin is a nucleoside antimetabolite with severe toxicity and teratogenic effects

Method used

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  • Neutralizing antibody against respiratory syncytial viruses and application thereof
  • Neutralizing antibody against respiratory syncytial viruses and application thereof
  • Neutralizing antibody against respiratory syncytial viruses and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Screening, Expression and Purification of Anti-Respiratory Syncytial Virus Neutralizing Antibodies

[0059] 1. Preparation and identification of RSV Pre-F protein

[0060] According to the gene sequence of the pre-fusion conformation (Pre-F) protein of the RSV A2 strain fusion (F) protein in the NCBI database, the expression plasmid of RSV Pre-F protein was constructed with the pcDNA3.3 expression vector after adding the His tag, and transfected into 293T After the cells were cultured, the supernatant expressed by the cells was collected, concentrated and then purified by a nickel column to obtain the RSV Pre-F protein, which was then detected and identified.

[0061] 1) detection of expressed RSV Pre-F protein

[0062] The expressed RSV Pre-F protein was detected by SDS-PAGE, and the result showed that its relative molecular weight was about 70kDa, which was consistent with the theoretical value.

[0063] The expressed RSV Pre-F protein was detected by Nati...

Embodiment 2

[0083] Example 2 Binding activity, in vitro micro-neutralizing activity and affinity experiments of neutralizing antibodies

[0084] 1. Antibody binding activity

[0085] Use the same ELISA method mentioned above to detect the binding activity of the expressed and purified antibody:

[0086] The RSV Pre-F protein was coated with carbonate coating buffer, overnight at 4°C. Wash with PBST buffer, add blocking solution to block for 2h at 37°C or overnight at 4°C. Use the blocking solution to dilute the RSV antibody sample proportionally, starting at 1 μg / well, and diluting 12 gradients. Positive controls are positive plasma sample stock solution and RSV monoclonal antibody, 100 μL per well. Negative control was negative plasma sample stock solution and irrelevant antibody (TRN006) 0.5 μg / mL, 100 μL per well, blank plus 100 μL blocking solution, and incubated at 37°C for 1 hour. Wash with PBST buffer, add 100 μL of Goat-Anti-Human IgG-Fab-HRP and Goat-Anti-Mouse IgG-Fab-HRP di...

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Abstract

The present invention relates to the field of medicine and immunology, and specifically discloses a neutralizing antibody against respiratory syncytial viruses and an application thereof. The neutralizing antibody comprises a heavy chain variable region of VH CDR1, VH CDR2 and VH CDR3 and a light chain variable region (or named as a variable light chain domain) of VL CDR1, VL CDR2 and VL CDR3; wherein amino acid sequences of the VH CDR1, VH CDR2 and VH CDR3 in the heavy chain variable region are shown in SEQ ID NO.1, 2 and 3, the amino acid sequences of the VL CDR1, VL CDR2 and VL CDR3 in thelight chain variable region are shown in SEQ ID NO.4, 5 and 6. Based on research results, the present invention also provides a nucleic acid molecule encoding the neutralizing antibody, and applications of the neutralizing antibody in preparing a pre-fusion protein product specifically binding the respiratory syncytial viruses, preparing respiratory syncytial virus vaccines, etc.

Description

technical field [0001] The invention relates to the fields of medicine and immunology, in particular to a neutralizing antibody against respiratory syncytial virus. Background technique [0002] Respiratory syncytial virus (RSV) is widely distributed throughout the world and is one of the most common viral pathogens causing lower respiratory illness (LRI) in infants, the elderly, and immunocompromised adults . Almost all children experience one or more infections by the age of 2 years, with the peak age of infection being 2 months to 8 months. RSV is the leading cause of lower respiratory tract infections in infants and young children, and the leading cause of respiratory hospitalization in young children. In hospitalized cases of infants and young children, 40% to 50% of bronchiolitis and 25% of pneumonia are caused by RSV infection. Moreover, many studies have also shown that severe infection in infants is a high risk factor for later asthma, and its severity far exceed...

Claims

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Application Information

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IPC IPC(8): C07K16/10C12N15/13G01N33/569A61K39/42A61P31/14
CPCA61K2039/505A61P31/14C07K16/1027C07K2317/565C07K2317/622C07K2317/76G01N33/56983G01N2333/135
Inventor 廖化新郑伟宏贾振兴王月明袁晓辉
Owner ZHUHAI TRINOMAB BIOTECHNOLOGY CO LTD
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