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Specific Probe Selection from Landscape Phage Display Library and Its Application in Enzyme-Linked Immunosorbent Assay of Free Prostate-Specific Antigen
Lang, Qiaolin1,2; Wang, Fei1,2,3; Yin, Long1,2,3; Liu, Mingjun4; Petrenko, Valery A.5; Liu, Aihua1,2,3
2014-03-04
发表期刊ANALYTICAL CHEMISTRY
卷号86期号:5页码:2767-2774
摘要Probes against targets can be selected from the landscape phage library f8/8, displaying random octapeptides on the pVIII coat protein of the phage fd-tet and demonstrating many excellent features including multivalency, stability, and high structural homogeneity. Prostate-specific antigen (PSA) is usually determined by immunoassay, by which antibodies are frequently used as the specific probes. Herein we found that more advanced probes against free prostate-specific antigen (f-PSA) can be screened from the landscape phage library. Four phage monoclones were selected and identified by the specificity array. One phage clone displaying the fusion peptide ERNSVSPS showed good specificity and affinity to f-PSA and was used as a PSA capture probe in a sandwich enzyme-linked immunosorbent assay (ELISA) array. An anti-human PSA monoclonal antibody (anti-PSA mAb) was used to recognize the captured antigen, followed by horseradish peroxidase-conjugated antibody (HRP-IgG) and o-phenylenediamine, which were successively added to develop plate color. The ELISA conditions such as effect of blocking agent, coating buffer pH, phage concentration, antigen incubation time, and anti-PSA mAb dilution for phage ELISA were optimized. On the basis of the optimal phage ELISA conditions, the absorbance taken at 492 nm on a microplate reader was linear with f-PSA concentration within 0.825 similar to 165 ng/mL with a low limit of detection of 0.16 ng/mL. Thus, the landscape phage is an attractive biomolecular probe in bioanalysis.
文章类型Article
WOS标题词Science & Technology ; Physical Sciences
DOI10.1021/ac404189k
关键词[WOS]RESOLVED IMMUNOFLUOROMETRIC ASSAY ; BIOCHEMICAL CHARACTERISTICS ; FILAMENTOUS BACTERIOPHAGE ; SALMONELLA-TYPHIMURIUM ; CANCER-DETECTION ; PEPTIDE LIBRARY ; MOLECULAR-FORMS ; TOTAL PSA ; SURFACE ; VIRUS
收录类别SCI
语种英语
WOS研究方向Chemistry
WOS类目Chemistry, Analytical
WOS记录号WOS:000332494100068
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文献类型期刊论文
条目标识符http://ir.qibebt.ac.cn/handle/337004/6191
专题生物传感技术团队(过去)
作者单位1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Lab Biosensing, Qingdao 266101, Peoples R China
2.Chinese Acad Sci, Key Lab Bioenergy, Qingdao 266101, Peoples R China
3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
4.Qingdao Univ, Affiliated Hosp, Coll Med, Dept Clin Lab, Qingdao 266003, Peoples R China
5.Auburn Univ, Dept Pathobiol, Auburn, AL 36849 USA
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Lang, Qiaolin,Wang, Fei,Yin, Long,et al. Specific Probe Selection from Landscape Phage Display Library and Its Application in Enzyme-Linked Immunosorbent Assay of Free Prostate-Specific Antigen[J]. ANALYTICAL CHEMISTRY,2014,86(5):2767-2774.
APA Lang, Qiaolin,Wang, Fei,Yin, Long,Liu, Mingjun,Petrenko, Valery A.,&Liu, Aihua.(2014).Specific Probe Selection from Landscape Phage Display Library and Its Application in Enzyme-Linked Immunosorbent Assay of Free Prostate-Specific Antigen.ANALYTICAL CHEMISTRY,86(5),2767-2774.
MLA Lang, Qiaolin,et al."Specific Probe Selection from Landscape Phage Display Library and Its Application in Enzyme-Linked Immunosorbent Assay of Free Prostate-Specific Antigen".ANALYTICAL CHEMISTRY 86.5(2014):2767-2774.
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