A protein chip approach for high-throughput antigen identification and characterization.
Proteomics research in humans and other eukaryotes demands a large number of high-quality mAbs. Here, we report a new approach to produce high-quality mAbs against human liver proteins using a combined force of high-throughput mAb production and protein microarrays. After immunizing mice with live cells from human livers, we isolated 54 hybridomas with binding activities to human cells and identified the corresponding antigens for five mAbs via screening on a protein microarray of 1058 unique human liver proteins. Finally, we demonstrated that using the five mAbs we could characterize the expression profiles of their corresponding antigens by using tissue microarrays. Among them, we discovered that eIF1A expressed only in normal liver tissues, not in hepatocellular carcinoma in humans.
Hu
Shaohui
S
Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China.
Li
Yu
Y
Liu
Guozhen
G
Song
Qifeng
Q
Wang
Li
L
Han
Yuning
Y
Zhang
Yang
Y
Song
Yali
Y
Yao
Xiying
X
Tao
Yong
Y
Zeng
Haipan
H
Yang
Huanming
H
Wang
Jian
J
Zhu
Heng
H
Chen
Zhi-Nan
ZN
Wu
Lin
L
D000328:Adult; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000941:Antigens / Q000032:analysis* / Q000276:immunology; D051926:Basigin / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D015415:Biomarkers / Q000032:analysis; D002352:Carrier Proteins / Q000032:analysis / Q000276:immunology; D049308:Dioxygenases; D015977:Eukaryotic Initiation Factor-1 / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D005260:Female; D005340:Fibrinogen; D006801:Humans; D007150:Immunohistochemistry; D008099:Liver / Q000737:chemistry / Q000378:metabolism; D008107:Liver Diseases / Q000378:metabolism; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology; D008565:Membrane Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D009363:Neoplasm Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D009687:Nuclear Proteins / Q000032:analysis / Q000276:immunology; D040081:Protein Array Analysis / Q000379:methods*; D011506:Proteins / Q000032:analysis / Q000276:immunology* / Q000378:metabolism; D015203:Reproducibility of Results; D014611:Vaccination
D000911:Antibodies, Monoclonal; D000941:Antigens; C497886:BSG protein, human; D015415:Biomarkers; D002352:Carrier Proteins; D015977:Eukaryotic Initiation Factor-1; C485894:FGL1 protein, human; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; C464279:ORMDL2 protein, human; D011506:Proteins; C026022:eukaryotic peptide initiation factor-1A; D051926:Basigin; D005340:Fibrinogen; D049308:Dioxygenases; C105340:PIR protein, human