A h7n9 virus specific recognition antibody p52h12 and detection kit
A P52H12 and detection kit technology, applied in the field of virus diagnosis, can solve problems such as loss of application value of humanized antibodies, loss of antibody affinity and specificity, difficulty in diagnosis of suspected cases, etc., to solve false positives, low cost, highly specific effect
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Embodiment 1H7
[0025] Example 1 Screening of H7N9 virus specific recognition antibody
[0026] (1) Construction of a phage display library of human Fab antibodies
[0027] The binding experiment was carried out with the convalescent patient serum after infection with H7N9 virus and SH02 HA, 17SF HA, human H5rHA monomer protein (SH02 MONO, 17SF MONO, human H5 MONO), and the experimental results are as follows image 3 As shown, it shows that the serum of the convalescent patients contains antibodies that can bind to H7 or H5 antigen, and BSA is used as a negative control, and there is no non-specific binding. The patient blood with specific binding was selected as the library sample.
[0028] Taking the patient's blood that can specifically bind to the above viral proteins as a sample, separate lymphocytes, extract total RNA, synthesize cDNA by reverse transcription, use cDNA as a template, and use specific primers to amplify the Fab fragment of the antibody. The resulting fragment was liga...
Embodiment 2
[0036] Example 2 Detection of properties of antibody P52H12
[0037] see figure 2 , is the result of affinity determination of P52H12 IgG antibody by ELISA, wherein ● indicates 17SFHA strain (A / Guangdong / 17SF003 / 2016), ■ indicates SH02 HA strain (A / Shanghai / 2 / 2013H7N9), ▲ indicates BSA negative control.
[0038] The results showed that the EC of P52H12 to the SH02 HA and 17SF HA proteins of two strains of H7N9 50 The values were 7.67nM and 34.4nM, respectively, with high affinity and certain cross-activity to the HA protein of H7 subtype; at the same time, the ELISA results showed that the antibody did not bind to the control BSA, and had no non-specificity. This antibody has no neutralizing inhibitory activity.
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