公开(公告)号：US09176136B2

公开(公告)日：2015-11-03

申请号：US14666869

申请日：2015-03-24

Applicant: OILCROPS RESEARCH INSTITUTE OF CHINESE ACADEMY OF AGRICULTURE SCIENCES

Inventor： Peiwu Li ,  Min Li ,  Qi Zhang ,  Zhaowei Zhang ,  Xiaoxia Ding

IPC: C07K16/14 ,  C07K16/00 ,  C07K14/38 ,  G01N33/569

CPC classification number: G01N33/56961 ,  C07K16/14 ,  C07K2317/92 ,  G01N2333/38 ,  G01N2469/10

Abstract: A hybridoma cell line ST03 having China Center for Type Culture Collection (CCTCC) accession number C2013187, a monoclonal antibody against aflatoxin biosynthetic precursor ST produced by the hybridoma cell line ST03, and the use of the monoclonal antibody. The hybridoma cell line ST03 can be used for preparing a high-titer monoclonal antibody against aflatoxin biosynthetic precursor ST, and the titer of mouse ascites antibody against aflatoxin biosynthetic precursor ST determined by enzyme linked immunosorbent assay (ELISA) can reach 6.4×105. The monoclonal antibody against aflatoxin biosynthetic precursor ST has high sensitivity, has 50% inhibiting concentration IC50 to aflatoxin biosynthetic precursor ST of 0.36 ng/mL, has no cross reaction with all of aflatoxin B1, aflatoxin B2, aflatoxin G1, and aflatoxin G2, and can be used for the content determination of aflatoxin biosynthetic precursor ST.

Abstract translation: 具有中国型号培养物保藏中心（CCTCC）登录号C2013187的杂交瘤细胞系ST03，由杂交瘤细胞系ST03产生的黄曲霉毒素生物合成前体ST的单克隆抗体，以及单克隆抗体的用途。 杂交瘤细胞系ST03可用于制备抗黄曲霉毒素生物合成前体ST的高滴度单克隆抗体，通过酶联免疫吸附试验（ELISA）测定的小鼠腹水抗黄曲霉毒素生物合成前体ST的滴度可达到6.4×105。 黄曲霉毒素生物合成前体ST的单克隆抗体具有较高的灵敏度，对黄曲霉毒素生物合成前体ST浓度IC50为0.36ng / mL，与黄曲霉毒素B1，黄曲霉毒素B2，黄曲霉毒素G1，黄曲霉毒素G2均无交叉反应， 可用于黄曲霉毒素生物合成前体ST的含量测定。

公开(公告)号：US20150276735A1

公开(公告)日：2015-10-01

申请号：US14667206

申请日：2015-03-24

Applicant: OILCROPS RESEARCH INSTITUTE OF CHINESE ACADEMY OF AGRICULTURE SCIENCES

Inventor： Peiwu Li ,  Ting He ,  Qi Zhang ,  Zhaowei Zhang ,  Xiaoxia Ding

IPC: G01N33/569 ,  C07K16/14

CPC classification number: G01N33/56961 ,  C07K16/14 ,  C07K2317/565 ,  C07K2317/569 ,  G01N2333/38 ,  G01N2469/10

Abstract: Aflatoxin M1 nanobody 2014AFM-G2 has the amino acid sequence of SEQ ID NO:7, and is encoded by the gene sequence of SEQ ID NO:8. The aflatoxin M1 nanobody 2014AFM-G2 obtained via screening has the properties of tolerance to organic reagents, tolerance to high temperature, tolerance to acids and bases and the like, and good stability. The aflatoxin M1 nanobody 2014AFM-G2 has 50% inhibiting concentration IC50 to aflatoxin M1 of 0.208 ng/mL, and has cross reaction rates with aflatoxin B1, B2, G1, G2 are 9.43%, 5.93%, 4.87% and 6.17%, respectively.

Abstract translation: 黄曲霉毒素M1纳米颗粒2014AFM-G2具有SEQ ID NO：7的氨基酸序列，由SEQ ID NO：8的基因序列编码。 通过筛选获得的黄曲霉毒素M1纳米粒子2014AFM-G2具有对有机试剂的耐受性，耐高温性，耐酸碱性等优良的稳定性。 黄曲霉毒素M1纳米粒子2014AFM-G2对黄曲霉毒素M1的IC50值为0.208 ng / mL，具有50％的黄曲霉毒素B1，B2，G1，G2的交叉反应率分别为9.43％，5.93％，4.87％和6.17％ 。

公开(公告)号：US09718836B2

公开(公告)日：2017-08-01

申请号：US14667341

申请日：2015-03-24

Applicant: OILCROPS RESEARCH INSTITUTE OF CHINESE ACADEMY OF AGRICULTURE SCIENCES

Inventor： Peiwu Li ,  Qi Zhang ,  Min Li ,  Fei Ma ,  Xiaoxia Ding

IPC: A61K39/02 ,  A61K39/00 ,  A61K39/385 ,  C07D493/14 ,  A61K31/335

CPC classification number: C07D493/14 ,  A61K31/335 ,  A61K39/0002

Abstract: An artificial antigen of aflatoxin biosynthetic precursor Sterigmatocystin (ST) and a method for preparing same. Firstly, hydroxyacetic acid is reacted with the double bound of the difuran ring in the aflatoxin biosynthetic precursor ST, yielding an aflatoxin biosynthetic precursor ST hapten with an active carboxymethoxy group. Secondly, a carboxyl group on the ST hapten is attached to an amino group on a carrier protein. At last, the artificial antigen of aflatoxin biosynthetic precursor ST is obtained by dialysis and lyophilize.

公开(公告)号：US09435776B2

公开(公告)日：2016-09-06

申请号：US14667378

申请日：2015-03-24

Applicant: OILCROPS RESEARCH INSTITUTE OF CHINESE ACADEMY OF AGRICULTURE SCIENCES

Inventor： Peiwu Li ,  Qi Zhang ,  Ting He ,  Zhaowei Zhang ,  Xiaoxia Ding

IPC: G01N33/543 ,  G01N1/40 ,  G01N30/02 ,  A61K51/10 ,  B01J20/281 ,  G01N30/00

CPC classification number: G01N30/48 ,  G01N33/54346 ,  G01N2030/486

Abstract: An aflatoxin M1 nanobody, an immunosorbent and an immunoaffinity column. The aflatoxin M1 nanobody 2014AFM-G2 has the amino acid sequence of SEQ ID NO:7, is encoded by the nucleic acid sequence of SEQ ID NO:8, has a 50% inhibiting concentration IC50 to aflatoxin M1 of 0.208 ng/mL, and has cross reaction rates with aflatoxins B1, B2, G1, and G2 of 9.43%, 5.93%, 4.87% and 6.17%, respectively. The immunosorbent includes a solid phase carrier and aflatoxin M1 nanobody 2014AFM-G2 coupled with the solid phase carrier. The immunoaffinity column is loaded with the aflatoxin M1 nanobody immunosorbent. It can be used for purifying and concentrating an extracting solution of a sample before loading to a machine for detection and the immunoaffinity column can be used repeatedly for many times.

Abstract translation: 黄曲霉毒素M1纳米体，免疫吸附剂和免疫亲和柱。 黄曲霉毒素M1纳米粒子2014AFM-G2具有由SEQ ID NO：8的核酸序列编码的SEQ ID NO：7的氨基酸序列，对黄曲霉毒素M1的IC50抑制率为0.208ng / mL，抑制浓度为50％， 黄曲霉毒素B1，B2，G1，G2的交叉反应率分别为9.43％，5.93％，4.87％和6.17％。 免疫吸附剂包括与固相载体偶联的固相载体和黄曲霉毒素M1纳米颗粒。 免疫亲和柱装载有黄曲霉毒素M1纳米体免疫吸附剂。 它可以用于在加载到机器进行检测之前将样品的提取溶液进行纯化和浓缩，并且免疫亲和柱可以重复使用多次。

公开(公告)号：US20150276734A1

公开(公告)日：2015-10-01

申请号：US14666869

申请日：2015-03-24

Applicant: OILCROPS RESEARCH INSTITUTE OF CHINESE ACADEMY OF AGRICULTURE SCIENCES

Inventor： Peiwu Li ,  Min Li ,  Qi Zhang ,  Zhaowei Zhang ,  Xiaoxia Ding

IPC: G01N33/569 ,  C07K16/14

CPC classification number: G01N33/56961 ,  C07K16/14 ,  C07K2317/92 ,  G01N2333/38 ,  G01N2469/10

Abstract: A hybridoma cell line ST03 having China Center for Type Culture Collection (CCTCC) accession number C2013187, a monoclonal antibody against aflatoxin biosynthetic precursor ST produced by the hybridoma cell line ST03, and the use of the monoclonal antibody. The hybridoma cell line ST03 can be used for preparing a high-titer monoclonal antibody against aflatoxin biosynthetic precursor ST, and the titer of mouse ascites antibody against aflatoxin biosynthetic precursor ST determined by enzyme linked immunosorbent assay (ELISA) can reach 6.4×105. The monoclonal antibody against aflatoxin biosynthetic precursor ST has high sensitivity, has 50% inhibiting concentration IC50 to aflatoxin biosynthetic precursor ST of 0.36 ng/mL, has no cross reaction with all of aflatoxin B1, aflatoxin B2, aflatoxin G1, and aflatoxin G2, and can be used for the content determination of aflatoxin biosynthetic precursor ST.

Abstract translation: 具有中国型号培养物保藏中心（CCTCC）登录号C2013187的杂交瘤细胞系ST03，由杂交瘤细胞系ST03产生的黄曲霉毒素生物合成前体ST的单克隆抗体，以及单克隆抗体的用途。 杂交瘤细胞系ST03可用于制备抗黄曲霉毒素生物合成前体ST的高滴度单克隆抗体，通过酶联免疫吸附试验（ELISA）测定的小鼠腹水抗黄曲霉毒素生物合成前体ST的滴度可达到6.4×105。 黄曲霉毒素生物合成前体ST的单克隆抗体具有较高的灵敏度，对黄曲霉毒素生物合成前体ST浓度IC50为0.36ng / mL，与黄曲霉毒素B1，黄曲霉毒素B2，黄曲霉毒素G1和黄曲霉毒素G2均无交叉反应， 可用于黄曲霉毒素生物合成前体ST的含量测定。

公开(公告)号：US09631010B2

公开(公告)日：2017-04-25

申请号：US15215056

申请日：2016-07-20

Applicant: OILCROPS RESEARCH INSTITUTE OF CHINESE ACADEMY OF AGRICULTURE SCIENCES

Inventor： Peiwu Li ,  Qi Zhang ,  Yanru Wang ,  Zhaowei Zhang ,  Xiaoxia Ding

IPC: G01N33/543 ,  G01N1/40 ,  G01N30/02 ,  C07K1/22 ,  C07K16/14 ,  G01N33/569

CPC classification number: C07K16/14 ,  C07K1/22 ,  C07K2317/22 ,  C07K2317/565 ,  C07K2317/569 ,  C07K2317/92 ,  C07K2317/94 ,  G01N33/56961 ,  G01N2333/38

Abstract: An aflatoxin nanobody immunoabsorbent and immunoaffinity column and preparation method and use thereof. The immunoabsorbent comprises a solid phase carrier and aflatoxin B1 nanobody 2014AFB-G15 coupled with the solid phase carrier. The 50% inhibiting concentration IC50 of aflatoxin B1 nanobody 2014AFB-G15 to aflatoxin B1 is 0.66 ng/mL, and the cross-reactivity of aflatoxin B1 nanobody 2014AFB-G15 to aflatoxins B2, G1, G2, and M1 are respectively 22.6%, 10.95%, 32.1% and 26%. The amino acid sequence of aflatoxin B1 nanobody 2014AFB-G15 is as depicted by SEQ ID NO: 7, and the coding gene sequence thereof is as depicted by SEQ ID NO: 8. The aflatoxin nanobody immunoaffinity column can be used for purification and concentration of sample extract prior to computer testing, and the immunoaffinity column can be reused repeatedly.

公开(公告)号：US20150276729A1

公开(公告)日：2015-10-01

申请号：US14667378

申请日：2015-03-24

Applicant: OILCROPS RESEARCH INSTITUTE OF CHINESE ACADEMY OF AGRICULTURE SCIENCES

Inventor： Peiwu Li ,  Qi Zhang ,  Ting He ,  Zhaowei Zhang ,  Xiaoxia Ding

IPC: G01N33/543 ,  B01J20/281

CPC classification number: G01N30/48 ,  G01N33/54346 ,  G01N2030/486

Abstract: An aflatoxin M1 nanobody, an immunosorbent and an immunoaffinity column. The aflatoxin M1 nanobody 2014AFM-G2 has the amino acid sequence of SEQ ID NO:7, is encoded by the nucleic acid sequence of SEQ IDNO:8, has a 50% inhibiting concentration IC50 to aflatoxin M1 of 0.208 ng/mL, and has cross reaction rates with aflatoxins B1, B2, G1, and G2 of 9.43%, 5.93%, 4.87% and 6.17%, respectively. The immunosorbent includes a solid phase carrier and aflatoxin M1 nanobody 2014AFM-G2 coupled with the solid phase carrier. The immunoaffinity column is loaded with the aflatoxin M1 nanobody immunosorbent. It can be used for purifying and concentrating an extracting solution of a sample before loading to a machine for detection and the immunoaffinity column can be used repeatedly for many times.

Abstract translation: 黄曲霉毒素M1纳米体，免疫吸附剂和免疫亲和柱。 黄曲霉毒素M1纳米粒子2014AFM-G2具有SEQ ID NO：7的氨基酸序列，由SEQ ID NO：8的核酸序列编码，对黄曲霉毒素M1的IC50值为0.208ng / mL，具有50％的抑制浓度，具有 黄曲霉毒素B1，B2，G1和G2的交叉反应率分别为9.43％，5.93％，4.87％和6.17％。 免疫吸附剂包括与固相载体偶联的固相载体和黄曲霉毒素M1纳米颗粒。 免疫亲和柱装载有黄曲霉毒素M1纳米体免疫吸附剂。 它可以用于在加载到机器进行检测之前将样品的提取溶液进行纯化和浓缩，并且免疫亲和柱可以重复使用多次。

公开(公告)号：US20150274846A1

公开(公告)日：2015-10-01

申请号：US14667341

申请日：2015-03-24

Applicant: OILCROPS RESEARCH INSTITUTE OF CHINESE ACADEMY OF AGRICULTURE SCIENCES

Inventor： Peiwu Li ,  Qi Zhang ,  Min Li ,  Fei Ma ,  Xiaoxia Ding

IPC: C07K19/00

CPC classification number: C07D493/14 ,  A61K31/335 ,  A61K39/0002

Abstract: An artificial antigen of aflatoxin biosynthetic precursor terigmatocystin (ST) and a method for preparing same. Firstly, hydroxyacetic acid is reacted with the double bound of the difuran ring in the aflatoxin biosynthetic precursor ST, yielding an aflatoxin biosynthetic precursor ST hapten with an active carboxymethoxy group. Secondly, a carboxyl group on the ST hapten is attached to an amino group on a carrier protein. At last, the artificial antigen of aflatoxin biosynthetic precursor ST is obtained by dialysis and lyophilize.

Abstract translation: 黄曲霉毒素生物合成前体特西他霉素（ST）的人造抗原及其制备方法。 首先，在黄曲霉毒素生物合成前体ST中，羟基乙酸与二呋喃环的双重界面反应，得到具有活性羧基甲氧基的黄曲霉毒素生物合成前体ST半抗原。 其次，ST半抗原上的羧基与载体蛋白上的氨基连接。 最后，通过透析和冻干获得黄曲霉毒素生物合成前体ST的人造抗原。

公开(公告)号：US09128089B1

公开(公告)日：2015-09-08

申请号：US14667206

申请日：2015-03-24

Applicant: OILCROPS RESEARCH INSTITUTE OF CHINESE ACADEMY OF AGRICULTURE SCIENCES

Inventor： Peiwu Li ,  Ting He ,  Qi Zhang ,  Zhaowei Zhang ,  Xiaoxia Ding

IPC: G01N33/53 ,  A61K51/10 ,  G01N33/569 ,  C07K16/14

CPC classification number: G01N33/56961 ,  C07K16/14 ,  C07K2317/565 ,  C07K2317/569 ,  G01N2333/38 ,  G01N2469/10

Abstract: Aflatoxin M1 nanobody 2014AFM-G2 has the amino acid sequence of SEQ ID NO:7, and is encoded by the gene sequence of SEQ ID NO:8. The aflatoxin M1 nanobody 2014AFM-G2 obtained via screening has the properties of tolerance to organic reagents, tolerance to high temperature, tolerance to acids and bases and the like, and good stability. The aflatoxin M1 nanobody 2014AFM-G2 has 50% inhibiting concentration IC50 to aflatoxin M1 of 0.208 ng/mL, and has cross reaction rates with aflatoxin B1, B2, G1, G2 are 9.43%, 5.93%, 4.87% and 6.17%, respectively.

Abstract translation: 黄曲霉毒素M1纳米颗粒2014AFM-G2具有SEQ ID NO：7的氨基酸序列，由SEQ ID NO：8的基因序列编码。 通过筛选获得的黄曲霉毒素M1纳米粒子2014AFM-G2具有对有机试剂的耐受性，耐高温性，耐酸碱性等优良的稳定性。 黄曲霉毒素M1纳米粒子2014AFM-G2对黄曲霉毒素M1的IC50值为0.208 ng / mL，具有50％的黄曲霉毒素B1，B2，G1，G2的交叉反应率分别为9.43％，5.93％，4.87％和6.17％ 。