Anti-prostate acid phosphatase antibody and application thereof

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

Active Publication Date: 2022-04-15
SHANGHAI HUIMMUTECH BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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However, there is still no method for efficiently

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  • Anti-prostate acid phosphatase antibody and application thereof
  • Anti-prostate acid phosphatase antibody and application thereof
  • Anti-prostate acid phosphatase antibody and application thereof

Examples

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preparation example Construction

[0190] The preparation method of the nucleic acid is a conventional preparation method in the art. Preferably, it includes the following steps: obtaining the nucleic acid molecule encoding the above-mentioned protein by gene cloning technology, or obtaining the 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 protein to provide a polynucleotide homologue. The homologue of the polynucleotide in the present invention can be prepared by replacing, deleting or adding one or more bases in the gene encoding the protein sequence within the scope of maintaining antibody activity.

[0192] carrier

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

[0194] The recombinant expression v...

Embodiment 1

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

[0253] The PAP-GM-CSF protein (refer to the inventor's patent application for the preparation method: preparation method of prostatic acid phosphatase / granulocyte-macrophage colony-stimulating factor, patent application number: 201810073775.X) was immunized with BalB / c mice several times , fused the splenocytes of immunized mice with SP2 / 0 myeloma cells to prepare hybridoma cells, and finally screened 9 monoclonal cell lines by ELISA method and monoclonalization method, and determined their ELISA binding curve and ED50 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: ...

Embodiment 2

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

[0277] experimental method:

[0278] The cell line highly expressing anti-PAP antibody obtained in Example 1 was revived, cultured, and transferred to finally obtain a culture supernatant highly expressing anti-PAP antibody.

[0279] The cell culture supernatant was collected by centrifugation, adjusted to pH using buffer A (20 mM PBS, pH 7.0), and loaded onto an affinity chromatography column equilibrated with buffer A. After loading the sample, use buffer A to wash the column until the A280 value and the conductance return to the baseline, use 100% buffer B (20mM citric acid, pH 2.7) for one-step elution, monitor the A280 value of the eluent, and collect the eluted fraction. The product was initially purified for anti-PAP-GM-CSF antibody.

[0280] Use buffer C (10mM citric acid, pH 5.5) to adjust the pH of the preliminary purified product to 5.0-6.0, load the sample onto the equilibrated cation ex...

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Abstract

The invention belongs to the field of biological medicine, and particularly provides a sequence of a prostate acid phosphatase (PAP) resisting antibody and application of the prostate acid phosphatase (PAP) resisting antibody. The anti-PAP antibody disclosed by the invention has relatively high affinity with PAP or a fusion protein of a prostate acid phosphatase-granulocyte-macrophage colony stimulating factor (PAP-GM-CSF), and can be used for purifying the PAP or the PAP-GM-CSF (Prostate acid phosphatase-granulocyte-macrophage colony stimulating factor), so that the anti-PAP antibody disclosed by the invention can be used for purifying the PAP or the PAP-GM-CSF (Prostate acid phosphatase-granulocyte-macrophage colony stimulating factor) and can be used for purifying the PAP or the PAP-GM-CSF (Prostate acid phosphatase-granulocyte-macrophage colony stimulating factor). Meanwhile, the kit can also be used for diagnosis and treatment of prostatic cancer.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to an anti-prostatic acid phosphatase antibody, its preparation method and application. Background technique [0002] Prostate cancer has been one of the most common cancers in men for decades, with new cases diagnosed each year in men and second only to lung cancer as the second leading cause of cancer death in men. In 2018, there were nearly 1.3 million new cases of prostate cancer diagnosed worldwide, and 359,000 prostate cancer-related deaths. From a global perspective, the incidence of prostate cancer in developed countries is higher than that in developing countries. With the rapid economic growth in developing countries, the incidence rate in 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 has increased year by year at an annual growth rate of 12.07%; in t...

Claims

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Application Information

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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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