Beijing Protein Innovation Co.

Beijing, China

Beijing Protein Innovation Co.

Beijing, China

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Huang H.,Peking University | Han Y.,Peking University | Gao J.,Peking University | Feng J.,Peking University | And 4 more authors.
Medical Oncology | Year: 2013

Gastric cancer is one of the most common human cancers and ranks the second in the global cancer-related mortality. The clinical outcome of patients with advanced gastric cancer (AGC) is markedly dependent on their response to the chemotherapy. Paclitaxel plus capecitabine, as a first-line regimen, is widely administrated in AGC patients, but more than a half of the patients have a poor response, possibly due to their resistance to the treatment. Therefore, it is important to identify potential responders to improve the efficacy of the chemotherapy. In the present study, we used an isobaric tag approach for relative and absolute quantification combined with ESI-QUAD-TOF/MS to identify potential predictive biomarkers for the chemotherapy. We found 211 serum proteins, and confirmed 17 candidates that were differentially present in the progression of disease (PD) group and the partial response (PR) group to the treatment of paclitaxel plus capecitabine. In further validation of the 17 candidates in the set of 12 PD and 12 PR AGC patients, we identified a higher level of AMBP (Alpha-1-Microglobulin/Bikunin Precursor) in the sera of PD patients than of the PR patients assayed by ELISA (9.13 ± 0.45 vs. 8.11 ± 0.26 μg/mL, p = 0.06) and by the Western blotting (relative gray value 396.4 ± 39.1 vs. 275.0 ± 34.76, p = 0.03), respectively. The receiver operating characteristics curve showed 75 % sensitivity and 75 % specificity of AMBP in AGC patients treated with the chemotherapy. Our data indicated that the high level of serum AMBP could predict the poor response of the AGC patients treated with the paclitaxel-capecitabine chemotherapy, which could be used as a potential biomarker to identify patients who would benefit from this chemotherapeutic regimen. © 2013 Springer Science+Business Media New York.


He L.-P.,CAS Institute of Physics | Liu S.,CAS Institute of Physics | Dai J.,CAS Institute of Physics | Wu L.,CAS Beijing Institute of Genomics | And 5 more authors.
Chinese Physics Letters | Year: 2015

Life science has a need for detection methods that are label-free and real-time. In this paper, we have selected staphylococcal protein A (SPA) and swine immunoglobulin G (IgG), and monitor the bindings between SPA and swine IgG with different concentrations, as well as the dissociations of SPA-swine IgG complex in different pH values of phosphate buffer by oblique-incidence reflectivity difference (OIRD) in a label-free and real-time fashion. We obtain the ON and OFF reaction dynamic curves corresponding to the bindings and dissociations of SPA and swine IgG. Through our analysis of the experimental results, we have been able to obtain the damping coefficients and the dissociation time of SPA and swine IgG for different pH values of the phosphate buffer. The results prove that the OIRD technique is a competing method for monitoring the dynamic processes of biomolecule interaction and achieving the quantitative information of reaction kinetics. © 2015 Chinese Physical Society and IOP Publishing Ltd.


Hu C.-J.,Peking Union Medical College | Song G.,CAS Beijing Institute of Genomics | Song G.,University of Chinese Academy of Sciences | Huang W.,CAS Beijing Institute of Genomics | And 14 more authors.
Molecular and Cellular Proteomics | Year: 2012

Primary biliary cirrhosis (PBC) is a chronic cholestatic liver disease of unknown etiology and is considered to be an autoimmune disease. Autoantibodies are important tools for accurate diagnosis of PBC. Here, we employed serum profiling analysis using a human proteome microarray composed of about 17,000 full-length unique proteins and identified 23 proteins that correlated with PBC. To validate these results, we fabricated a PBC-focused microarray with 21 of these newly identified candidates and nine additional known PBC antigens. By screening the PBC microarrays with additional cohorts of 191 PBC patients and 321 controls (43 autoimmune hepatitis, 55 hepatitis B virus, 31 hepatitis C virus, 48 rheumatoid arthritis, 45 systematic lupus erythematosus, 49 systemic sclerosis, and 50 healthy), six proteins were confirmed as novel PBC autoantigens with high sensitivities and specificities, including hexokinase-1 (isoforms I and II), Kelch-like protein 7, Kelch-like protein 12, zinc finger and BTB domain-containing protein 2, and eukaryotic translation initiation factor 2C, subunit 1. To facilitate clinical diagnosis, we developed ELISA for Kelch-like protein 12 and zinc finger and BTB domain-containing protein 2 and tested large cohorts (297 PBC and 637 control sera) to confirm the sensitivities and specificities observed in the microarray-based assays. In conclusion, our research showed that a strategy using high content protein microarray combined with a smaller but more focused protein microarray can effectively identify and validate novel PBC-specific autoantigens and has the capacity to be translated to clinical diagnosis by means of an ELISAbased method. © 2012 by The American Society for Biochemistry and Molecular Biology, Inc.


Huang W.,CAS Beijing Institute of Genomics | Huang W.,University of Chinese Academy of Sciences | Hu C.,Peking Union Medical College | Zeng H.,Beijing Protein Innovation Co. | And 8 more authors.
Biochemical and Biophysical Research Communications | Year: 2012

Systemic lupus erythematosus (SLE) is a systemic autoimmune disease affecting many organs. Many autoantibodies have been associated with the disease, but either in low specificity or low sensitivity of detection. In an aim to screen for better autoantibodies, we profiled the autoantibody repertoire in sera from 30 SLE patients versus 30 healthy controls using a protein microarray containing 5011 non-redundant human proteins, and identified four candidates. We then selected CLIC2 for further verification by ELISA in an extended cohort including 110 SLE, 121 non-AD, 118 RA, 117 SSc, and 105 pSS patients. The positive rate of anti-CLIC2 was 28.18% in SLE patients, significantly higher than those in non-AD, RA, and SSc patients. The presence of anti-CLIC2 in SLE had positive correlation with disease activity in terms of SLEDAI score and several indexes (p< 0.05). © 2012 Elsevier Inc.


Song G.,Peking Union Medical College | Song G.,CAS Beijing Institute of Genomics | Hu C.,Peking Union Medical College | Zhu H.,Beijing Protein Innovation Ltd. Co | And 7 more authors.
BMC Gastroenterology | Year: 2013

Background: Primary biliary cirrhosis (PBC) is a liver specific chronic disease with unclear pathogenesis, especially for the early stage molecular events. The mitochondrion is a multi-functional organelle associated with various diseases including PBC. The purpose of this study was to discover the alterations in the mitochondria proteome using an early stage PBC mouse model for revealing the possible pathogenesis mechanisms in the early stages of PBC.Methods: Mouse model of early stage of PBC was constructed by consecutive administration of poly I:C. Mitochondria of mouse models and controls were purified and comparative proteomics was performed by iTRAQ technology. Then, differentially expressed proteins were validated by western blotting.Results: In total 354 proteins that satisfied the criteria for comparative proteomics study were identified. Of them, nine proteins were downregulated and 20 were up-regulated in liver mitochondria of PBC mouse model. Most differentially expressed proteins are associated with oxidation-reduction and lipid metabolism, and some are involved in the biosynthesis of steroid hormone and primary bile acid. Interestingly, four proteins (HCDH, CPT I, DECR, ECHDC2) involved in the fatty acid beta-oxidation were all upregulated.Conclusions: iTRAQ is a powerful tool for comparative proteomics study of PBC mouse model and differentially expressed proteins in mitochondria proteome of PBC mouse model provide insights for the pathogenesis mechanism at early stage of PBC. © 2013 Song et al.; licensee BioMed Central Ltd.


Chen X.,Chinese Academy of Sciences | Chen X.,Guangxi University | Zhu H.,Chinese Academy of Sciences | Hu C.,Guangxi University | And 9 more authors.
Reproduction | Year: 2014

Cryodamage is a major problem in semen cryopreservation, causing changes in the levels of proteins that influence the function and motility of spermatozoa. In this study, protein samples prepared from fresh and frozen-thawed boar spermatozoa were compared using the isobaric tags for relative and absolute quantification (iTRAQ) labeling technique coupled to 2D LC-MS/MS analysis. A total of 41 differentially expressed proteins were identified and quantified, including 35 proteins that were present at higher levels and six proteins that were present at lower levels in frozen-thawed spermatozoa by at least a mean of 1.79-fold (P<0.05). On classifying into ten distinct categories using bioinformatic analysis, most of the 41 differentially expressed proteins were found to be closely relevant to sperm premature capacitation, adhesions, energy supply, and sperm-oocyte binding and fusion. The expression of four of these proteins, SOD1, TPI1, ODF2, and AKAP3, was verified by western blot analysis.We propose that alterations in these identified proteins affect the quality of cryopreserved semen and ultimately lower its fertilizing capacity. This is the first study to compare protein levels in fresh and frozen- thawed spermatozoa using the iTRAQ technology. Our preliminary results provide an overview of the molecular mechanisms of cryodamage in frozen-thawed spermatozoa and theoretical guidance to improve the cryopreservation of boar semen. © 2014 Society for Reproduction and Fertility.


Bai H.,Hebei Academy of Agriculture and Forestry science | Bai H.,CAS Institute of Genetics and Developmental Biology | Bai H.,CAS Beijing Institute of Genomics | Lan J.P.,Agricultural University of Hebei | And 11 more authors.
Plant Biology | Year: 2012

Rice Xa21 gene encodes a receptor-like kinase that confers broad-spectrum resistance against Xanthomonas oryzae pv. oryzae (Xoo). Recently, a number of genes involved in the Xa21-mediated disease resistance pathway have been identified. Based on our previous data and the literature, we chose 16 candidate proteins and made corresponding antibodies. Using Western blotting, we systematically investigated the expression profile of the proteins in Xa21-mediated disease resistance response. We found nine proteins with altered expression. We further compared their expression in resistance, susceptible and mock responses, and found that GST expression was up-regulated during the resistance process, indicating GST is a positive regulator in resistance responses. ATPsB expression was down-regulated during both the resistance and susceptible response processes, although it was higher in the resistance response than that in the susceptible response. The total amount of MYB, GAPDH, CatB, Trx and NB-ARC proteins was lower in the resistance than in the susceptible response, but their abundance per unit bacteria in the resistance response was still higher than in the susceptible response, suggesting that these proteins might be positive regulators in the resistance response. In addition, expression of another ERF was induced by inoculation with bacterial blight pathogen, and expression of Zf-LSD1 was activated by wounding stress alone. Interestingly, most proteins showed similar altered expression patterns in the resistance and susceptible responses, but differed to some extents, implying that both responses might share common molecular mechanisms. This study revealed evidence of resistance-related protein expression, providing a foundation for better understanding of their functions. © 2012 German Botanical Society and The Royal Botanical Society of the Netherlands.


Lan J.-P.,Agricultural University of Hebei | Wu P.-C.,Beijing Protein Innovation Co. | Guo M.-C.,Agricultural University of Hebei | Shi J.-N.,Agricultural University of Hebei | And 10 more authors.
Progress in Biochemistry and Biophysics | Year: 2015

The npt II encoding neomycin phosphotransferase æ, is one of the most commonly used selection marker in transgenic research. In this study, recombinant NPT II protein was expressed in E. coli and monoclonal antibodies were generated. The inheritance, temp-spatial profiling, abundance and subcellular localization properties were investigated using established Western blot (WB) analysis. The results showed that NPT II protein in 0.25%of a single rice grain (about 0.025mg) was detectable, the expression of NPT II protein is consistent with dominant inheritance law, positive seeds and homozygous lines can be identified in transgenic T1 generation, furthermore, the abundance of NPT II protein can provide clues for the identification of homozygous individual in T1seedlings. Quantitative analyses shows that NPTæprotein accounts for about 0.08% of total protein in the leaves at seedling stage. The expression profile of NPT II protein in transgenic rice shows that the abundance of NPT II driven by CaMV-35S promoter in the leaves and roots at seedling and adult stage are higher than that at tillering and booting stage. NPT II was detectable at different stages in rice root, stem, leaf, panicle, flower and seed, however, the abundance of NPT II in tillers, node, panicle axis and anther tissues is lower than others. In addition, the NPT II protein is mainly localized in the soluble portion of cytoplasm. Taken together, an applicable immunoblot method was established and the expression properties of NPT II protein in transgenic rice were demonstrated.


Rong R.-J.,Agricultural University of Hebei | Wu P.-C.,Beijing Protein Innovation Co. | Lan J.-P.,Agricultural University of Hebei | Wei H.-F.,Beijing Protein Innovation Co. | And 11 more authors.
Journal of Integrative Agriculture | Year: 2016

Phosphomannose isomerase (PMI) encoding gene manA is a desirable selective marker in transgenic research. Understanding of its expression patterns in transgenic plant and establishing highly sensitive detection method based on immunoassay have great impacts on the application of PMI. In this study, PMI-specific monoclonal antibodies were generated using recombinant protein as immunogen, and could be used in Western blot to detect as little as 0.5 ng His-tagged PMI protein or rice expressed PMI protein in sample accounted for 0.4% of single rice grain (about 0.08 mg). PMI protein driven by CaMV-35S promoter was detected in dozens of tested tissues, including root, stem, leaf, panicle, and seed at all developmental stages during rice growing, and PMI protein accounted for about 0.036% of total protein in the leaves at seedling stage. The established method potentially can be used to monitor PMI protein in rice grains. © 2016 Chinese Academy of Agricultural Sciences.

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