Controlled antibody immobilization onto immunoanalytical platforms by synthetic peptide

Jung, Yongwon; Kang, Hyo Jin; Lee, Jeong Min; Jung, Sun Ok; Yun, Wan Soo; Chung, Sang J.; Chung, Bong Hyun

doi:10.1016/j.ab.2007.10.022

Cited by 88 publications

(52 citation statements)

References 19 publications

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“…(b) Lateral flow assay of a target analyte using magnetic labels. Conjugate pad Test line Control line Absorbent pad ported, and the use of protein A or G was shown to be effective for oriented immobilization of antibodies [13][14][15][16] .…”

Section: Construction Of a Magnetic Label By Oriented Immobilization mentioning

confidence: 99%

Increase in the detection sensitivity of a lateral flow assay for a cardiac marker by oriented immobilization of antibody

et al. 2011

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Early detection of human cardiac markers is regarded as a gold standard for diagnosis of acute myocardial infarction (AMI). Here we demonstrate the increase in the detection sensitivity of a lateral flow assay for a cardiac marker troponin I (cTnI) by oriented immobilization of antibodies on magnetic beads. For the construction of sensitive magnetic labels, orientated-immobilization of anti-troponin I antibodies on magnetic beads was attempted using protein G. Magnetic beads were first conjugated with protein G followed by immobilization of anti-cTnI antibodies. The factors affecting the detection sensitivity in the lateral flow assay system were investigated. The lateral flow assay of cTnI was carried out using the magnetic beads and a Giant magnetoresistive (GMR) sensor at the optimized conditions, and compared the results with using a conventional random immobilization method. Magnetic labels with oriented immobilization of antibodies enabled the detection of cTnI up to 0.01 ng/mL, showing significantly increased sensitivity compared with conventional immobilization method. This result clearly shows that the magnetic labels with oriented immobilization of antibodies can find wide applications in the lateral flow assays of target analytes.

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Section: Construction Of a Magnetic Label By Oriented Immobilization mentioning

confidence: 99%

Increase in the detection sensitivity of a lateral flow assay for a cardiac marker by oriented immobilization of antibody

et al. 2011

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“…Similarly, a synthesized Fc-binding cyclic peptide containing 13 amino acids was modified with a soluble poly (ethylene glycol) linker in the N-terminus and was uniformly coupled to glass slides to bind the Fc domain of antibodies (Jung et al, 2008). To achieve an appropriate orientation (Gering, 2002), the preparation method is based on a condensation reaction between an aldehyde group on the Fc portion of the antibody and a hydrazide group on the modified glass surface.…”

Section: Protein A/g-mediated Bioaffinity Immobilizationmentioning

confidence: 99%

Technological Development of Antibody Immobilization for Optical Immunoassays: Progress and Prospects

Wang

et al. 2014

Critical Reviews in Analytical Chemistry

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The high sensitivity of optical detection techniques and the highly specific reactions between antibodies and antigens mean that optical immunoassays have attracted much interest in the fields of protein, hormone, drug, and microorganism detection, without the need for complex separation and extraction steps. The immobilization of an antibody on a solid support is a crucial step for optical immunoassays. This review surveys the latest advances in current antibody immobilization techniques in detail, including physical adsorption, covalent attachment, bioaffinity immobilization, and some recently developed methods. Furthermore, specific consideration is given to oriented immobilization, which may improve the homogeneous surface covering the accessibility of the active site and surface coverage, and the analytical performance of immunoassays. Finally, new perspectives for antibody immobilization techniques in optical immunoassays are outlined.

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“…Immobilization of proteins through self-assembly onto solid surfaces with high affinity and material selectivity is an essential component in building up protein chips, protein microarrays, and biosensors (Bailey et al, 2007;Jung et al, 2008;Kwon et al, 2006;Rusmini et al, 2007;Tomizaki et al, 2005;Watzke et al, 2006). Although, there are many methods for protein immobilization on to solid surfaces, site-specific protein immobilization has been increasingly used to maintain protein conformation and retained biological activity on selected solid surfaces (Kumada et al, 2006;Kwon et al, 2006;Park et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

Directed self‐immobilization of alkaline phosphatase on micro‐patterned substrates via genetically fused metal‐binding peptide

Kacar

Zin

et al. 2009

Biotech & Bioengineering

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Current biotechnological applications such as biosensors, protein arrays, and microchips require oriented immobilization of enzymes. The characteristics of recognition, self-assembly and ease of genetic manipulation make inorganic binding peptides an ideal molecular tool for site-specific enzyme immobilization. Herein, we demonstrate the utilization of gold binding peptide (GBP1) as a molecular linker genetically fused to alkaline phosphatase (AP) and immobilized on gold substrate. Multiple tandem repeats (n = 5, 6, 7, 9) of gold binding peptide were fused to N-terminus of AP (nGBP1-AP) and the enzymes were expressed in E. coli cells. The binding and enzymatic activities of the bi-functional fusion constructs were analyzed using quartz crystal microbalance spectroscopy and biochemical assays. Among the multiple-repeat constructs, 5GBP1-AP displayed the best bi-functional activity and, therefore, was chosen for self-immobilization studies. Adsorption and assembly properties of the fusion enzyme, 5GBP1-AP, were studied via surface plasmon resonance spectroscopy and atomic force microscopy. We demonstrated self-immobilization of the bi-functional enzyme on micro-patterned substrates where genetically linked 5GBP1-AP displayed higher enzymatic activity per area compared to that of AP. Our results demonstrate the promising use of inorganic binding peptides as site-specific molecular linkers for oriented enzyme immobilization with retained activity. Directed assembly of proteins on solids using genetically fused specific inorganic-binding peptides has a potential utility in a wide range of biosensing and bioconversion processes.

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Controlled antibody immobilization onto immunoanalytical platforms by synthetic peptide

Cited by 88 publications

References 19 publications

Increase in the detection sensitivity of a lateral flow assay for a cardiac marker by oriented immobilization of antibody

Increase in the detection sensitivity of a lateral flow assay for a cardiac marker by oriented immobilization of antibody

Technological Development of Antibody Immobilization for Optical Immunoassays: Progress and Prospects

Directed self‐immobilization of alkaline phosphatase on micro‐patterned substrates via genetically fused metal‐binding peptide

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