Artron BioResearch Inc.

Burnaby, Canada

Artron BioResearch Inc.

Burnaby, Canada
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Cao H.,Chongqing Medical University | Xiang T.,Artron BioResearch Inc. | Zhang C.,Chongqing Medical University | Yang H.,Chongqing Medical University | And 3 more authors.
Biomedicine and Pharmacotherapy | Year: 2016

Background/aim To investigate the therapeutic effect of MDA7 combined with apoptin targeted attenuated Salmonella typhimurium vector SL7207/pBud-VP3 on gastric cancer cells. Materials and methods MDA7 was inserted into pBud-VP3 using molecular cloning technology to obtain the eukaryotic expression plasmid pBud-VP3-MDA7 and it was transformed into attenuated Salmonella typhimurium SL7207 by high voltage electroporation to obtain SL7207/pBud-VP3-MDA7. Mice bearing a sarcoma of gastric cancer cells were treated with SL7207/pBud-VP3-MDA7 and the growth-suppressing effect was assessed by measurement of tumor volume. Western blot was used to identify the MDA7 expression products. IL-6, INF-γ, TNF-α and caspase-3, VEGF in tumor tissue were detected by RT-PCR and immunohistochemistry. Results SL7207/pBud-VP3-MDA7 was successfully constructed and expression of the protein MDA7 was identified in tumor tissue. SL7207/pBud-VP3-MDA7 significantly caused tumor inhibition and regression (p < 0.05). The level of expression of cytokines IL-6, INF-γ, TNF-α in tumor tissue was significantly higher than in the other groups (p < 0.05). The expression of caspase-3 was up-regulated and VEGF was down-regulated (p < 0.05). Conclusion This study shows that SL7207/pBud-VP3-MDA7 has inhibitory effect on the growth of gastric cancer cells. The mechanism involved is related to the promotion of tumor apoptosis, immunity regulation and inhibition of tumor blood vessels. © 2016 Elsevier Masson SAS


PubMed | Jinan Kangbo Biotechnology. 2711 Ying Xiu Road, Artron BioResearch Inc., Chongqing Medical University and Shandong Jiaotong Hospital
Type: Journal Article | Journal: Clinical biochemistry | Year: 2015

Helicobacter pylori stool antigen (HpSA) test is a convenient and reliable non-invasive diagnosis for H. pylori infection. The aim of this study is to develop an immunochromatographic testing device for rapid detection of HpSA among Chinese patients.Monoclonal antibodies (McAb) targeting H. pylori were developed by conventional methods and paired by sandwich ELISA. The lateral flow device (LFD) was prepared using the selected McAb pair. A total of 867 clinically separated bacterial strains, including 56H. pylori strains, were employed to test the sensitivity and specificity. Subsequently, the LFD was used to test 1200 human fecal samples, with a commercial HpSA testing device as comparison.Two McAb pairs targeting H. pylori, DF2a/EE10b and IH10b/EE10b, were developed and proven to be of high specificity and sensitivity. After testing the cultures of 56 clinically separated H. pylori strains, the final LFD product was prepared using the mixture of DF2a and IH10b as capture antibodies and EE10b as the detection antibody. The testing threshold for H. pylori culture was 1.0 10(4)cfu/mL. The sensitivity and specificity were both 100% for the 867 tested bacterial cultures. The testing results of 1200 fecal samples showed that the positive and negative agreement rates between the homemade LFD and the commercial testing device were 99.75% and 99.87%, respectively.Our homemade HpSA LFD can be a very promising testing device for rapid diagnosis and epidemic screening of H. pylori infection among Chinese patients.


PubMed | Artron BioResearch Inc., Chongqing Medical University and Shandong Jiaotong Hospital
Type: | Journal: Protein expression and purification | Year: 2016

Alanine aminotransferase (ALT) has been used as a sensitive marker for liver injury in people and in preclinical toxicity studies. But measurement of ALT isoenzymes, ALT1 and ALT2, was reported to be of more diagnostic value. The aim of this study is to develop an ideal pair of anti-ALT1 monoclonal antibodies (MAbs) of high specificity and affinity, and subsequently prepare a Immunochromatographic lateral flow device (LFD) for rapid test of ALT1 in human serums.The complete coding sequence of ALT1 gene (1500 bp) was cloned from human hepatoma G2 cells (HepG2) and inserted into the expression vector pET-32a(+). ALT1 recombinant protein was routinely prepared by E. coli BL21 (DE3) expression and Ni(2+) affinity purification. Balb/c mice were immunized with purified ALT1 and the splenocytes were fused with Sp2/0 myeloma cells. The positive clones, verified by indirect enzyme-linked immunosorbent assay (ELISA) using purified ALT1, were subcloned to single clones by limiting dilution process. A MAb pair was selected from the obtained MAbs according the sandwich ELISA pairing results and then used for lateral flow device (LFD) production. After evaluation of the sensitivity and specificity, the LFD strips were employed to test human serum samples with known ALT activity levels.ALT1 recombinant protein was expectedly prepared by expression and purification. A total of 8 stable clones that produced antibodies specifically recognizing ALT1 protein were developed. After sandwich ELISA pairing, an ideal pair of anti-ALT1 MAbs, designated as BD7 and DG3, were selected and proved to be of high specificity, titer and affinity. Based on the MAb pair, LFD strips specifically for ALT1 rapid test were subsequently prepared. The detection threshold of the LFD strips was 12 U/L. No cross reaction was found.The ALT1 LFD with high sensitivity and specificity was successfully developed. It is valuable for testing ALT1 protein in human sera and can be a beneficial complement for traditional ALT test.


Xiang T.,Chongqing Medical University | Jiang Z.,Chongqing Medical University | Zheng J.,Artron BioResearch Inc. | Lo C.,Artron BioResearch Inc. | And 5 more authors.
International Journal of Molecular Medicine | Year: 2012

The objective of this study was to screen for antigens of the hepatitis C virus (HCV) to establish a new double antibody sandwich-lateral flow immunoassay (DAS-LFIA) method for testing the presence of anti-HCV antibodies in human serum or plasma. A series of different recombinant HCV proteins in Escherichia coli cells were constructed, expressed, purified and the new DAS-LFIA strip was developed. The sensitivity and specificity of new the DAS-LFIA strip were evaluated by detecting 23 HCV-positive sera, a set of quality control references for anti-HCV detection that contain known amounts of anti-HCV antibodies, and 8 HCV-negative sera. A total of 300 clinical serum samples was examined by both the new DAS-LFIA strip and enzyme-linked immunosorbent assay (ELISA). Data were analyzed using SPSS 11.5 software. The sensitivity and specificity of the new DAS-LFIA strip were 100%. The lowest test line of the HCV DAS-LFIA strips was 2 NCU/ml. Additionally, the concordance between the new DAS-LFIA strip and ELISA methods was 94.33%. In conclusion, our new testing method is rapid, simple, sensitive and specifically detects the presence of anti-HCV antibodies in human serum or plasma. Therefore, it may be used for monitoring HCV.


Chen W.,Chongqing Medical University | Zhang J.,Artron BioResearch Inc. | Lu G.,Artron BioResearch Inc. | Yuan Z.,Chongqing Medical University | And 6 more authors.
Clinical Biochemistry | Year: 2014

Objectives: Cholera is an acute malignant infectious disease caused by the bacteria Vibrio cholerae leading to severe dehydrating diarrhea and vomiting, even high rates of mortality in some cases. However, the prevention of the epidemic disease is achievable if proper sanitation practices are followed, provided the accurate and prompt diagnosis of each prevalent serotype in cholera epidemic. The current gold standard of bacterial culture is inadequate for rapid diagnosis. Our aim is to develop an immunochromatographic test format for O1 serotype Ogawa diagnosis and provide the need for better epidemic prevention and early response. Design and methods: The monoclonal antibodies were raised in conventional method and subsequently screened for a match pair. A variety of related and unrelated bacteria strains recruited were employed to test their sensitivity, specificity etc. by indirect ELISA. The human fecal samples were used to test the final lateral-flow device product to satisfy the measurement requirement. Results: A new monoclonal antibody (McAb) pair, named IXiao3G6 and IXiao1D9, was generated, which is specifically against V. cholerae O1 serotype Ogawa. Additionally, we developed an immunochromatographic lateral flow device (LFD) using this McAb pair for the highly specific and rapid (5min) detection of Ogawa. Conclusions: Our product has advantages of simplicity and precision, and can benefit the scene and elementary medical institutions. © 2013 The Canadian Society of Clinical Chemists.


Xu G.,Chongqing Medical University | Zou W.-Q.,Chongqing Medical University | Du S.-J.,Chongqing Medical University | Wu M.-J.,Chongqing Medical University | And 2 more authors.
Life Sciences | Year: 2016

Aims Prostate cancer (PCa) is one of the most common cancers in men in the world. Advanced PCa, especially castration-resistant PCa (CRPC), is difficult to cure. There is an urgent need to develop novel agents for CPRC. Dihydroartemisinin (DHA) is a semisynthetic derivative of artemisinin and is a well-known antimalarial drug. DHA has been documented to be a potential anticancer agent for PCa. However, the mechanisms underlying the anticancer activity of DHA are still unknown. Main methods Proteomics analysis based on iTRAQ technology was performed to determine the protein profile changes in human prostate cancer PC3 cells treated by DHA, and apoptosis was detected by flow cytometry and transmission electron microscopy. Key findings DHA induced obvious apoptosis in PC3 cells. Using iTRAQ technology, we found 86 differentially expressed proteins linked to the cytotoxicity of DHA in PC3 cells. Gene ontology analysis showed the differentially expressed proteins were mainly associated with the protein synthesis and translation. Protein interaction network analysis and KEGG pathway analysis revealed altered aminoacyl-tRNA biosynthesis and metabolic pathways. Moreover, one candidate protein, heat shock protein HSP70 (HSPA1A), was identified by western blot analysis. Significance Our results indicate that multiple mechanisms involved in the anticancer activity of DHA in PC3 cells. Decreased HSP70 expression may have an important role in DHA-induced apoptosis in PC3 cells. Our data also provide novel insights into the anticancer mechanisms of DHA. © 2016


PubMed | Artron BioResearch Inc. and Chongqing Medical University
Type: | Journal: Life sciences | Year: 2016

Prostate cancer (PCa) is one of the most common cancers in men in the world. Advanced PCa, especially castration-resistant PCa (CRPC), is difficult to cure. There is an urgent need to develop novel agents for CPRC. Dihydroartemisinin (DHA) is a semisynthetic derivative of artemisinin and is a well-known antimalarial drug. DHA has been documented to be a potential anticancer agent for PCa. However, the mechanisms underlying the anticancer activity of DHA are still unknown.Proteomics analysis based on iTRAQ technology was performed to determine the protein profile changes in human prostate cancer PC3 cells treated by DHA, and apoptosis was detected by flow cytometry and transmission electron microscopy.DHA induced obvious apoptosis in PC3 cells. Using iTRAQ technology, we found 86 differentially expressed proteins linked to the cytotoxicity of DHA in PC3 cells. Gene ontology analysis showed the differentially expressed proteins were mainly associated with the protein synthesis and translation. Protein interaction network analysis and KEGG pathway analysis revealed altered aminoacyl-tRNA biosynthesis and metabolic pathways. Moreover, one candidate protein, heat shock protein HSP70 (HSPA1A), was identified by western blot analysis.Our results indicate that multiple mechanisms involved in the anticancer activity of DHA in PC3 cells. Decreased HSP70 expression may have an important role in DHA-induced apoptosis in PC3 cells. Our data also provide novel insights into the anticancer mechanisms of DHA.


PubMed | and Ji Nan Kangbo Biotechnology, Artron BioResearch Inc., Chinese Academy of Agricultural Sciences and Chongqing Medical University
Type: Evaluation Studies | Journal: Journal of agricultural and food chemistry | Year: 2015

Phytase is a phosphohydrolase considered highly specific for the degradation of phytate to release bound phosphorus for animal consumption and aid in the reduction of environmental nutrient loading. New sources of phytase have been sought that are economically and efficiently productive including the construction of genetically modified (GM) phytase products designed to bypass the costs associated with feed processing. Four monoclonal antibodies (EH10a, FA7, AF9a, and CC1) raised against recombinant Aspergillus niger phyA2 were used to develop a highly specific and sensitive immunochromatographic lateral flow device for rapid detection of transgenic phytase, such as in GM corn. Antibodies sequentially paired and tested along lateral flow strips showed that the EH10a-FA7 antibody pair was able to detect the recombinant yeast-phytase at 5 ng/mL, whereas the AF9a-CC1 antibody pair to GM phytase corn was able to detect at 2 ng/mL. Concurrent to this development, evidence was revealed which suggests that antibody binding sites may be glycosylated.


PubMed | Artron BioResearch Inc. and Chongqing Medical University
Type: | Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie | Year: 2016

To investigate the therapeutic effect of MDA7 combined with apoptin targeted attenuated Salmonella typhimurium vector SL7207/pBud-VP3 on gastric cancer cells.MDA7 was inserted into pBud-VP3 using molecular cloning technology to obtain the eukaryotic expression plasmid pBud-VP3-MDA7 and it was transformed into attenuated Salmonella typhimurium SL7207 by high voltage electroporation to obtain SL7207/pBud-VP3-MDA7. Mice bearing a sarcoma of gastric cancer cells were treated with SL7207/pBud-VP3-MDA7 and the growth-suppressing effect was assessed by measurement of tumor volume. Western blot was used to identify the MDA7 expression products. IL-6, INF-, TNF- and caspase-3, VEGF in tumor tissue were detected by RT-PCR and immunohistochemistry.SL7207/pBud-VP3-MDA7 was successfully constructed and expression of the protein MDA7 was identified in tumor tissue. SL7207/pBud-VP3-MDA7 significantly caused tumor inhibition and regression (p<0.05). The level of expression of cytokines IL-6, INF-, TNF- in tumor tissue was significantly higher than in the other groups (p<0.05). The expression of caspase-3 was up-regulated and VEGF was down-regulated (p<0.05).This study shows that SL7207/pBud-VP3-MDA7 has inhibitory effect on the growth of gastric cancer cells. The mechanism involved is related to the promotion of tumor apoptosis, immunity regulation and inhibition of tumor blood vessels.


PubMed | Artron BioResearch Inc. and Chongqing Medical University
Type: Journal Article | Journal: Clinical biochemistry | Year: 2014

Cholera is an acute malignant infectious disease caused by the bacteria Vibrio cholerae leading to severe dehydrating diarrhea and vomiting, even high rates of mortality in some cases. However, the prevention of the epidemic disease is achievable if proper sanitation practices are followed, provided the accurate and prompt diagnosis of each prevalent serotype in cholera epidemic. The current gold standard of bacterial culture is inadequate for rapid diagnosis. Our aim is to develop an immunochromatographic test format for O1 serotype Ogawa diagnosis and provide the need for better epidemic prevention and early response.The monoclonal antibodies were raised in conventional method and subsequently screened for a match pair. A variety of related and unrelated bacteria strains recruited were employed to test their sensitivity, specificity etc. by indirect ELISA. The human fecal samples were used to test the final lateral-flow device product to satisfy the measurement requirement.A new monoclonal antibody (McAb) pair, named IXiaoG and IXiaoD, was generated, which is specifically against V. cholerae O1 serotype Ogawa. Additionally, we developed an immunochromatographic lateral flow device (LFD) using this McAb pair for the highly specific and rapid (5 min) detection of Ogawa.Our product has advantages of simplicity and precision, and can benefit the scene and elementary medical institutions.

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