TY - JOUR
T1 - Biological function analysis of monoclonal antibodies against human granulins in vitro using U251 cells as a model
AU - Li, Yanqing
AU - Li, Ya
AU - Ye, Mingfu
AU - Wang, Dongyang
AU - Zhao, Junli
AU - Sun, Xiaohong
AU - Mao, Qinwen
AU - Xia, Haibin
N1 - Funding Information:
This work was supported by the Fundamental Research Funds for the Central Universities ( GK2015030 ), research grants to H.X. from the National Natural Science Foundation of China (No. 81272543 , No. 81471772 ) and the Natural Science Foundation of Shaanxi Province, China (No. 2014JM4113 ).
Publisher Copyright:
© 2016 Elsevier Inc.
PY - 2017/2/1
Y1 - 2017/2/1
N2 - Progranulin (PGRN), a highly glycosylated, secreted 593 amino acid precursor protein, is a multifunctional molecule that is critical for early embryogenesis, wound repair, inflammatory and tumorigenesis. PGRN can be proteolytically cleaved into seven cysteine-rich granulin (Grn) peptides: G, F, B, A, C, D and E. Both PGRN and its constituent Grn peptides have been implicated in a wide variety of biological activities. However, their functions are far from clear, and the lack of granulin domain-specific antibodies has hindered the progress of the functional study of PGRN and Grns. Monoclonal antibodies against GrnB, GrnA, GrnC and GrnF have been previously developed by our laboratory. In this study, we generated monoclonal antibodies (MAbs) against GrnD, GrnG and GrnE by using recombinant proteins HSA-GrnG, HSA-GrnD and HSA-GrnE as immunogens, and characterized them by indirect ELISA, Western blot and immunocytochemistry. Furthermore, the neutralizing activities of the MAbs against seven Grns were tested in vitro using the U251 cell line. This full antibody panel of MAbs against seven Grns will be a valuable tool for elucidating the biological roles of PGRN and Grns in different physiopathological processes, which will further promote the development of PGRN-based clinical diagnosis and therapy.
AB - Progranulin (PGRN), a highly glycosylated, secreted 593 amino acid precursor protein, is a multifunctional molecule that is critical for early embryogenesis, wound repair, inflammatory and tumorigenesis. PGRN can be proteolytically cleaved into seven cysteine-rich granulin (Grn) peptides: G, F, B, A, C, D and E. Both PGRN and its constituent Grn peptides have been implicated in a wide variety of biological activities. However, their functions are far from clear, and the lack of granulin domain-specific antibodies has hindered the progress of the functional study of PGRN and Grns. Monoclonal antibodies against GrnB, GrnA, GrnC and GrnF have been previously developed by our laboratory. In this study, we generated monoclonal antibodies (MAbs) against GrnD, GrnG and GrnE by using recombinant proteins HSA-GrnG, HSA-GrnD and HSA-GrnE as immunogens, and characterized them by indirect ELISA, Western blot and immunocytochemistry. Furthermore, the neutralizing activities of the MAbs against seven Grns were tested in vitro using the U251 cell line. This full antibody panel of MAbs against seven Grns will be a valuable tool for elucidating the biological roles of PGRN and Grns in different physiopathological processes, which will further promote the development of PGRN-based clinical diagnosis and therapy.
KW - Eukaryotic expression
KW - Hybridoma
KW - Monoclonal antibody
KW - Progranulin
KW - Prokaryotic expression
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U2 - 10.1016/j.pep.2016.09.019
DO - 10.1016/j.pep.2016.09.019
M3 - Article
C2 - 27693922
AN - SCOPUS:84991434209
SN - 1046-5928
VL - 130
SP - 55
EP - 62
JO - Protein Expression and Purification
JF - Protein Expression and Purification
ER -