Production and Characterization of a Monoclonal Antibody Against GRAM Domain-Containing Protein 1A

Song, Xiuli; Wang, Shihao; Gu, Bin; Hou, Qiannv; Liu, Yajuan; Zhang, Ming

doi:10.1089/mab.2014.0001

Cited by 3 publications

(4 citation statements)

References 14 publications

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“…GRAMD1A is widely expressed in the nucleus and cytoplasm of embryonic stem cells, cancer cell lines, and ectodermal, mesodermal, and endodermal tissues, and its function has not been fully explored [ 8 ]. In previous studies, GRAMD1A was found to accumulate at autophagosome initiation sites, influence cholesterol distribution during starvation, and participate in autophagosome biogenesis [ 24 ].…”

Section: Discussionmentioning

confidence: 99%

“…GRAM domain-containing protein 1A (GRAMD1A), originally identified in human embryonic stem cells, is 724 amino acids in length and contains a GRAM structural domain [ 8 ]; it is thought to be an intracellular protein-binding or lipid-binding signalling domain and may play an important role in membrane-associated processes [ 9 ]. GRAMD1A is found to be expressed in a variety of human cancer tissues, including the digestive, urinary, and reproductive systems [ 8 ]. In addition, GRAMD1A was found to promote the expansion of hepatocellular carcinoma stem cells, the growth of hepatocellular carcinoma, and resistance to chemotherapy via STAT5 [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

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GRAMD1A Is a Biomarker of Kidney Renal Clear Cell Carcinoma and Is Associated with Immune Infiltration in the Tumour Microenvironment

Liu

Zhang

et al. 2022

Disease Markers

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Background. GRAM structural domain-containing protein 1A (GRAMD1A) is upregulated in a variety of human cancer tissues and is closely associated with tumourigenesis and progression. Methods. Patient RNA-sequencing data and clinicopathological information were obtained from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets. The expression of GRAMD1A in kidney cancer cell lines and KIRC patients was analysed by quantitative polymerase chain reaction (qPCR). Receiver Operator Characteristic (ROC) curves, nomograms, Kaplan-Meier analysis, forest plots, and COX analysis were used to assess the diagnostic and prognostic value of GRAMD1A in KIRC, and gene set enrichment analysis (GSEA) was used to explore its potential signalling pathways. In addition, the Sangerbox website, Kaplan-Meier plotter database, and TISIDB and TIMER databases were used to further analyse the correlation of GRAMD1A with microsatellite instability (MSI), tumour mutational burden (TMB), immune checkpoint genes, and tumour-infiltrating lymphocytes (TILs). Results. GRAMD1A was significantly highly expressed in KIRC and associated with shorter overall survival and relapse-free survival ( P < 0.05 ). The AUC value of the ROC curve to identify KIRC and normal renal tissues was 0.942. Forest plot and COX analysis visualized that GRAMD1A could be an independent prognostic factor in KIRC patients ( P < 0.01 ), and nomograms to determine the overall survival (OS) of KIRC patients also showed good efficacy (C-index: 0.776). Moreover, Spearman correlation analysis showed a positive correlation between GRAMD1A and MSI, TMB ( P < 0.01 ). On the other hand, GRAMD1A was also found to be closely associated with immune checkpoint genes. Meanwhile, patients with KIRC with high GRAMD1A expression had a relatively low hazard ratio (HR) of death when B lymphocytes, natural killer T cells, CD4+ T lymphocytes, CD8+ T lymphocytes, and macrophages were enriched in the tumour microenvironment (TME), and a greater HR of death when regulatory T lymphocytes with tumour-specific immunosuppressive effects were significantly enriched. Last, GSEA shows that GRAMD1A is closely associated with the regulation of energy metabolism in KIRC. Conclusions. GRAMD1A is a promising diagnostic and prognostic biomarker for patients with KIRC, and its biological function correlates to some extent with immune infiltration in TME.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

GRAMD1A Is a Biomarker of Kidney Renal Clear Cell Carcinoma and Is Associated with Immune Infiltration in the Tumour Microenvironment

Liu

Zhang

et al. 2022

Disease Markers

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“…The class and subclass of the isotypes of the purified antibody were determined by using a mouse monoclonal antibody isotyping kit. Measurement of monoclonal antibody affinity (Ka) was carried out according to the procedure described by Jiang et al [5] . An indirect competitive ELISA (icELISA) standard curve was developed based on the mAbs.…”

Section: Characterization Of Monoclonal Antibodiesmentioning

confidence: 99%

Preparation and Immunological Traits of Monoclonal Antibody against Sarafloxacin

Fan

Jiang

2012

AMR

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Through cell fusion technology, five hybridoma lines of sarafloxacin (SAR), named S1-B2, S2-C6, S2-E7, S3-C5, and S3-E5, were identified and their corresponding mAbs were of the IgG1 isotype with a k light chain. The Kaffs of all mAbs were between 2.8 and 4.6×109 L/mol. The titers and IC50 values of purified ascite fluids were in the range of 0.512–2.56×106 and 0.32–0.48 ng/mL, respectively. The performances of S1-B2 and S2-C6 were more consistent in the stability experiments. Based on the S1-B2 hybridoma, an icELISA method was developed. The dynamic range was from 0.004 to 18 ng/mL, with a detection limit for the assay and IC50 values of 0.002 and 0.32 ng/mL, respectively. Therefore, the establishment of these hybridomas may provide an alternative method for the detection of SAR residues in food-original animals.

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“…IL-6/Jak2/Stat3 pathway promotes HCC growth, chemoresistance and the self-renewal of HCC stem cells91011. GRAMD1A first finds in human embryonic stem cell, and expresses in ectoderm, mesoderm and endoderm tissues and many tumor cells12, its function hasn’t been explored. Here, we studied the role of GRAMD1A in the prognosis, initiation and progression of HCC, we analyzed the correlation between GRAMD1A levels and many clinicopathological features, and examined whether GRAMD1A could function as a prognostic factor for HCC patients.…”

mentioning

confidence: 99%

GRAM domain-containing protein 1A (GRAMD1A) promotes the expansion of hepatocellular carcinoma stem cell and hepatocellular carcinoma growth through STAT5

Wang

Zhao

et al. 2016

Sci Rep

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Hepatocellular carcinoma (HCC) is the leading cause for cancer death worldwide, new prognostic factors and targets are critical for HCC treatment. Here, we found GRAMD1A was upregulated in HCC tissues, patients with high GRAMD1A levels had poor outcome, statistical analyses found GRAMD1A expression was positively correlated with pathologic differentiation and survival or mortality. It was an unfavorable prognostic factor for HCC patients. Functional analyses revealed GRAMD1A contributed to the self-renewal of HCC stem cells, resistance to chemotherapy and tumor growth of HCC determined by hepatosphere formation assay, side population (SP) analysis, TUNEL assay, soft agar growth ability assay and tumor growth model in vivo. Mechanism analyses found signal transducer and activator of transcription 5 (STAT5) was the target of GRAMD1A, GRAMD1A regulated the target genes of STAT5 and the transcriptional activity of STAT5. Inhibition of STAT5 in indicated HCC cells overexpressing GRAMD1A suppressed the effects of GRAMD1A on the self-renewal of HCC stem cell, resistance to chemotherapy and tumor growth, suggesting GRAMD1A promoted the self-renewal of HCC stem cells and the development of HCC by increasing STAT5 level. GRAMD1A might be a useful biomarker and target for HCC.

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Production and Characterization of a Monoclonal Antibody Against GRAM Domain-Containing Protein 1A

Cited by 3 publications

References 14 publications

GRAMD1A Is a Biomarker of Kidney Renal Clear Cell Carcinoma and Is Associated with Immune Infiltration in the Tumour Microenvironment

GRAMD1A Is a Biomarker of Kidney Renal Clear Cell Carcinoma and Is Associated with Immune Infiltration in the Tumour Microenvironment

Preparation and Immunological Traits of Monoclonal Antibody against Sarafloxacin

GRAM domain-containing protein 1A (GRAMD1A) promotes the expansion of hepatocellular carcinoma stem cell and hepatocellular carcinoma growth through STAT5

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