阅读说明：本技术 识别链格孢菌的单克隆抗体及其杂交瘤细胞株AaC5 (Monoclonal antibody capable of recognizing alternaria and hybridoma cell strain AaC5 thereof ) 是由 赵伟春 徐云飞 李吉二 祁依佳 于 2019-11-29 设计创作，主要内容包括：本发明公开了一种识别链格孢菌的单克隆抗体及其杂交瘤细胞株,所述识别链格孢菌的单克隆抗体,由保藏编号为：CCTCC No：C2019249的杂交瘤细胞株分泌产生；所述杂交瘤细胞株命名为杂交瘤细胞株AaC5,于2019年10月17日保藏于中国典型培养物保藏中心(CCTCC),保藏编号为：CCTCC No：C2019249。该单克隆抗体被用于由链格孢菌侵染引发的动植物病害的鉴定、动态监测以及链格孢菌的生物学研究,通过BaLb/c小鼠腹腔注射该细胞株,即可获得大量的该单克隆抗体,与多克隆抗体相比,具有纯度高,专一性强,重复性好等优点。(The invention discloses a monoclonal antibody for identifying alternaria and a hybridoma cell strain thereof, wherein the monoclonal antibody for identifying alternaria is prepared from the following components in parts by weight: CCTCC No: c2019249 hybridoma cell strain secretion; the hybridoma cell strain is named as hybridoma cell strain AaC5, is preserved in China Center for Type Culture Collection (CCTCC) in 2019, 10 and 17 months, and has the preservation number as follows: CCTCC No: C2019249. the monoclonal antibody is used for identification and dynamic monitoring of animal and plant diseases caused by Alternaria alternata infection and biological research of Alternaria alternata, and a large amount of monoclonal antibody can be obtained by injecting the cell strain into the abdominal cavity of a Balb/c mouse.)

1. A monoclonal antibody recognizing Alternaria alternata represented by the deposit number: CCTCC No: c2019249 is secreted and produced.

2. A hybridoma cell strain is named as hybridoma cell strain AaC5, is preserved in China Center for Type Culture Collection (CCTCC) in 2019, 10 months and 17 days, and has the preservation number as follows: CCTCC No: C2019249.

Technical Field

The invention belongs to the field of animals, and particularly relates to a monoclonal antibody for identifying alternaria alternata and a hybridoma cell strain AaC5 thereof.

Background

Alternaria alternata, a fungus of Alternaria of the subdivision Deuteromycotina, is one of the pathogenic fungi of fritillaria melasma. The pathogenic bacteria can be lost in soil with the hypha to live through the winter, and the fritillaria is infected again in the next year. Alternaria is also the pathogenic bacterium of alternaria leaf spot of many crops, causing serious losses to agricultural production every year. The alternaria fungus is of various types, and the colony, hypha and spore forms of related species are similar, so that the alternaria fungus is difficult to distinguish by means of the characteristics. The monoclonal antibody (monoclonal antibody) combined with enzyme-linked immunosorbent assay (ELISA) for Alternaria alternata disclosed by the invention has the characteristics of high sensitivity and strong specificity, and is suitable for detecting Alternaria alternata in fields in large quantities, so that a foundation is laid for the identification and biological research of Alternaria alternata by applying the antibody and dynamically monitoring the occurrence of diseases infected by Alternaria alternata, such as Fritillaria thunbergii black spot and the like.

Disclosure of Invention

The invention provides a monoclonal antibody for identifying alternaria, which is prepared from the following components in percentage by weight: CCTCC No: c2019249 is secreted and produced.

The invention also provides a hybridoma cell strain which is named as hybridoma cell strain AaC5 and is preserved in China Center for Type Culture Collection (CCTCC) in 2019, 10 months and 17 days, wherein the preservation numbers are as follows: CCTCC No: C2019249.

the invention has the following beneficial effects:

(1) the monoclonal antibody has strong specificity: the monoclonal antibody has strong reaction with Alternaria alternata antigen, does not react with Alternaria alternata, Fusarium oxysporum, Fusarium solani, Fusarium equiseti, Fusarium semitectum, Botrytis cinerea, Phoma niponicum and phomopsis longipes antigen, and can be used for detecting Alternaria alternata and thunberg fritillary black spot caused by the Alternaria alternata.

(2) The detection sensitivity of the monoclonal antibody is high: the indirect ELISA detection result shows that the detection sensitivity of the monoclonal antibody to the antigen prepared from the alternaria alternate hypha and spore is 24.42ng/mL (namely 2.442ng per well), and the monoclonal antibody has good development and application prospects.

(3) The monoclonal antibody is IgG 3.

(4) The monoclonal antibody-bound target protein is single: the monoclonal antibody only binds to an antigen protein of about 66kDa of Alternaria alternata.

(5) The detection sensitivity of the fritillaria thunbergii extracting solution on the monoclonal antibody is not influenced: the monoclonal antibody has no cross reaction to the thunberg fritillary bulb protein extracting solution; after the thunberg fritillary bulb protein extracting solution is added into the antigen prepared from the alternaria alternate hypha and the spores, the detection sensitivity of the thunberg fritillary bulb extracting solution on the monoclonal antibody is not influenced, the content is 24.42ng/mL (namely 2.442ng per hole), and the method has good development and application prospects.

Drawings

Potency assay of FIG. 1AaC 5;

FIG. 2AaC5 reaction with different fungal antigens;

the detection sensitivity assay of FIG. 3AaC 5;

type and subclass identification of FIG. 4AaC 5;

FIG. 5AaC5 detection of bound target protein;

FIG. 6 shows the effect of Bulbus Fritillariae Thunbergii extract on the detection sensitivity of AaC 5.

Detailed Description

The invention is further explained below with reference to the figures and examples.