Anti-prostatic acid phosphatase antibody and use thereof

A technology of acid phosphatase and prostate, applied in the field of biomedicine

Active Publication Date: 2022-07-01
SHANGHAI HUIMMUTECH BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still no method for efficiently producing this fusion protein PAP-GM-CSF in the art

Method used

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  • Anti-prostatic acid phosphatase antibody and use thereof
  • Anti-prostatic acid phosphatase antibody and use thereof
  • Anti-prostatic acid phosphatase antibody and use thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0190] The preparation method of the nucleic acid is a conventional preparation method in the field, preferably, comprising the following steps: obtaining a nucleic acid molecule encoding the above-mentioned protein by gene cloning technology, or obtaining a nucleic acid molecule encoding the above-mentioned protein by artificial full sequence synthesis .

[0191] Those skilled in the art know that substitutions, deletions, alterations, insertions or additions can be appropriately introduced into the base sequence encoding the amino acid sequence of the above-mentioned proteins to provide a homolog of a polynucleotide. Homologs of the polynucleotides of the present invention can be prepared by substituting, deleting or adding one or more bases of the gene encoding the protein sequence within the range that maintains the activity of the antibody.

[0192] carrier

[0193] The present invention also provides a recombinant expression vector comprising the nucleic acid.

[0194]...

Embodiment 1

[0252] Example 1: Preparation and screening of anti-PAP antibody monoclonal cell lines

[0253] BalB / c mice were immunized multiple times with PAP-GM-CSF protein (refer to the patent application of the inventor for the preparation method: preparation method of prostatic acid phosphatase / granular-macrophage colony-stimulating factor, patent application number: 201810073775.X) , spleen cells of immunized mice and SP2 / 0 myeloma cells were fused to prepare hybridoma cells. By ELISA and monoclonalization, 9 monoclonal cell lines were finally screened, and their ELISA binding curves and ED50 were determined. value.

[0254] (1) Materials and instruments

[0255] 1. Mice: BalB / c

[0256] 2. Myeloma cells: SP2 / 0 (ATCC)

[0257] 3. Immunogen: PAP-GM-CSF protein (Shanghai Huidun Biotechnology Co., Ltd., 1mg / ml)

[0258] 4. Cell growth medium: 1640+10%FBS(Gibco)+OPI Media Supplement(Sigma)

[0259] 5. Detection antibody: M-HRP-anti-mouse IgG

[0260] 6. Cell fusion instrument: BTX ...

Embodiment 2

[0276] Example 2: Preparation of the PAP antibody of the present invention

[0277] experimental method:

[0278] The cell line with high expression of anti-PAP antibody obtained in Example 1 was then recovered, cultured and transferred to finally obtain a culture supernatant with high expression of anti-PAP antibody.

[0279] Cell culture supernatants were collected by centrifugation, pH adjusted with buffer A (20 mM PBS, pH 7.0), and loaded onto an affinity chromatography column that had been equilibrated with buffer A. After loading, use buffer A to wash the column to the A280 value and conductance back to the baseline, use 100% buffer B (20mM citric acid, pH 2.7) to elute in one step, monitor the eluent A280 value, and collect the elution fractions. The product was initially purified for anti-PAP-GM-CSF antibody.

[0280] The pH of the preliminary purified product was adjusted to 5.0-6.0 with buffer C (10 mM citric acid, pH 5.5) and loaded onto an equilibrated cation exc...

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Abstract

The present invention belongs to the field of biomedicine, and more particularly, the present invention provides a sequence of an anti-prostatic acid phosphatase (PAP) antibody and use thereof. The anti-PAP antibody disclosed in the present invention has higher levels of PAP or prostatic acid phosphatase-granulocyte-macrophage colony-stimulating factor fusion protein (Fusion Protein of Prostatic acid phosphatase-Granulocyte-macrophage colony-stimulating factor, PAP-GM-CSF). The affinity can be used for the purification of PAP or PAP-GM-CSF. At the same time, it can also be used for the diagnosis and treatment of prostate cancer.

Description

technical field [0001] The invention belongs to the field of biomedicine, and particularly relates to an anti-prostatic acid phosphatase antibody, a preparation method and application thereof. Background technique [0002] Prostate cancer has been one of the most common cancers in men for decades, with new cases in men every year after lung cancer and the second leading cause of cancer death in men. In 2018, there were nearly 1.3 million newly diagnosed cases of prostate cancer worldwide, and 359,000 prostate cancer-related deaths. Globally, the incidence of prostate cancer in developed countries is higher than in developing countries. With the rapid economic growth of developing countries, the incidence of developing countries has also increased rapidly in recent years. According to statistics, from 1998 to 2008, the number of prostate cancer patients in my country increased year by year at an annual growth rate of 12.07%; in the past 10 years, the number of male prostate...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/40C07K19/00C07K1/22C07K1/18C12N15/13C12N5/10B01J20/29B01D15/38B01J20/30A61K39/395A61K47/68A61P35/00G01N33/574G01N33/573
CPCC07K16/40C07K14/535C12N9/16B01J20/291B01D15/3804A61K47/6871A61K47/6801A61P35/00G01N33/57434G01N33/57484G01N33/573C12Y301/03002C07K2317/56C07K2317/565C07K2317/567C07K2317/52C07K2317/24C07K2317/92C07K2319/02C07K2319/21C07K2319/41C07K2319/42C07K2319/43A61K2039/505G01N2333/916
Inventor 吴昊蒋应明陈国友
Owner SHANGHAI HUIMMUTECH BIOTECHNOLOGY CO LTD
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