A Duplexed Microsphere-Based Fluorescent Immunoassay

Szurdoki, Ferenc; Michael, Karri L.; Walt, David R.

doi:10.1006/abio.2001.5041

Cited by 58 publications

(35 citation statements)

References 46 publications

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“…While the limit of specific detection is considerably higher than that of more mature assays, the speed and multiplexing offered by this approach are promising. Other approaches, based on microfluidic arrays and photosensors, have also been reported and show promising improvements in sensitivity with no loss in specificity (81,82). The main advantages offered by these experimental systems is that in addition to automation, multiplexing, and throughput, they offer a quantitative assessment of the agent present, detecting in some instances fewer than 50 copies of target agent.…”

Section: Future Trendsmentioning

confidence: 99%

Immunological Methods for Detection and Identification of Infectious Disease and Biological Warfare Agents

Peruski

2003

Clin Vaccine Immunol

196

119

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NATURE OF THE PROBLEMBW agents. The release of a biological weapon (BW) agent by a terrorist group or military force would likely be silent and undetectable or nearly so. As shown by anthrax attack during the fall of 2001 in the eastern United States, patients would begin appearing at hospitals and clinics within several days of exposure, most presenting with nonspecific flu-like symptoms. The first days of the outbreak might not even cause undue concern. However, depending on the type of agent and the method of dispersal, the public healthcare system would rapidly be stretched to capacity and beyond.The qualities that make a good BW agent are its relationship between aerosolization, infectivity, or toxicity and the amount of agent required to produce an effect (48). In addition, criteria such as environmental stability, ease of production, disease severity, and communicability determine which agents are the most likely to be utilized. For maximum effect, an optimal agent should be highly lethal and easily produced in large quantities and have limited options for preventive or prophylactic treatment. Given that the respiratory route is the most effective for most BW agents, stability in an aerosol form and the capability to be readily dispersed also in an aerosol (1-to 10-m particle size) are necessary. When potential agents are reviewed for these characteristics, Bacillus anthracis (anthrax) and variola major virus (smallpox) are considered to have the greatest potential for mass casualties and civil disruption. Also high on a prospective list of agents are botulinum neurotoxins, Yersinia pestis, and Francisella tularensis (48,91,92). Lower on the prospective list are Burkholderia pseudomallei and Burkholderia mallei, Rickettsia sp., Coxiella burnetii, Venezuelan equine encephalitis virus, Marburg and Ebola viruses, and influenza viruses (48,63,91,92).Emerging infectious disease agents. In addition to diseases caused by intentional epidemics, there are several emerging infectious diseases (ID) with the potential for significant public health consequences, including dengue fever, West Nile fever, and Rift Valley fever as well as the recent reemergence of malaria in the eastern United States (48,63,91,92). As with BW agents, emerging ID agents may be directly transmissible or vector borne (63). A complex interplay of factors can influence disease emergence, including genetic variation, environmental changes, and population pressures. Further compounding this already complicated situation, are the estimated 600 million international tourists annually, many with the potential to the spread disease globally in a matter of hours (63). Clearly, the challenges facing modern clinical microbiologists and immunologists are daunting enough without the added difficulties posed by the intentional release of BW/ID agents! Because of the threat posed by both BW and emerging or reemerging ID agents, there is a need to rapidly identify such agents in the clinical setting in order to treat the individuals at risk and to improve p...

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Section: Future Trendsmentioning

confidence: 99%

Immunological Methods for Detection and Identification of Infectious Disease and Biological Warfare Agents

Peruski

2003

Clin Vaccine Immunol

196

119

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“…Images were acquired with a Photometrics 300 CCD camera operated at Ϫ40°C interfaced with an Apple G3 computer and the Metamorph imaging software from Universal Imaging Corporation (Downingtown, PA). Measurement of the fluorescence intensity of cells was as described previously with modifications (21). Briefly, images of a minimum of three separate fields containing an average of 90 cells were acquired and stored as electronic files.…”

Section: Wisp-1mentioning

confidence: 99%

WISP-1 Binds to Decorin and Biglycan

Desnoyers¹,

Arnott²,

Pennica³

2001

Journal of Biological Chemistry

110

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Wnt-1-induced secreted protein 1 (WISP-1) is a member of the CCN (connective tissue growth factor, Cyr61, NOV) family of growth factors. Structural and experimental evidence suggests that CCN family member activities are modulated by their interaction with sulfated glycoconjugates. To elucidate the mechanism of action for WISP-1, we characterized the specificity of its tissue and cellular interaction and identified binding factors. WISP-1 binding was restricted to the stroma of colon tumors and to cells with a fibroblastic phenotype. By using a solid phase assay, we showed that human skin fibroblast conditioned media contained WISP-1 binding factors. Competitive inhibition with different glycosaminoglycans and treatment with glycosaminoglycan lyases and proteases demonstrated that binding to the conditioned media was mediated by dermatan sulfate proteoglycans. Mass spectrometric analysis identified the isolated binding factors as decorin and biglycan. Decorin and biglycan interacted directly with WISP-1 and inhibited its binding to components in the conditioned media. Similarly, WISP-1 interaction with human skin fibroblasts was inhibited by dermatan sulfate, decorin, and biglycan or by treatment of the cell surface with dermatan sulfate-specific lyases. Together these results demonstrate that decorin and biglycan are WISP-1 binding factors that can mediate and modulate its interaction with the surface of fibroblasts. We propose that this specific interaction plays a role in the regulation of WISP-1 function.

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“…Future work will focus primarily on these two aspects. In addition to planar, two-dimensional constructs, this general approach would also be suitable for microbead systems [29] and should also find potential application in lab-on-a-chip [30] formats. We have demonstrated the feasibility of this approach with mRNA-protein fusions, but these methods should also be applicable to proteins that have been chemically derivatized with nucleic acids.…”

Section: Discussionmentioning

confidence: 99%

Generating addressable protein microarrays with PROfusion™ covalent mRNA-protein fusion technology

Weng¹,

Gu²,

Hammond³

et al. 2002

PROTEOMICS

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An mRNA-protein fusion consists of a polypeptide covalently linked to its corresponding mRNA. These species, prepared individually or en masse by in vitro translation with a modified mRNA conjugate (the PROfusion process), link phenotype to genotype and enable powerful directed evolution schemes. We have exploited the informational content of the nucleic acid component of the mRNA-protein fusion to create an addressable protein microarray that self-assembles via hybridization to surface-bound DNA capture probes. The nucleic acid component not only directs the mRNA-protein fusion to the proper coordinate of the microarray, but also positions the protein in a uniform orientation. We demonstrate the feasibility of this protein chip concept with several mRNA-protein fusions, each possessing a unique peptide epitope sequence. These addressable proteins could be visualized on the microarray both by autoradiography and highly specific monoclonal antibody binding. The anchoring of the protein to the chip surface is surprisingly robust, and the system is sensitive enough to detect sub-attomole quantities of displayed protein without signal amplification. Such protein arrays should be useful for functional screening in massively parallel formats, as well as other applications involving immobilized peptides and proteins.

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A Duplexed Microsphere-Based Fluorescent Immunoassay

Cited by 58 publications

References 46 publications

Immunological Methods for Detection and Identification of Infectious Disease and Biological Warfare Agents

Immunological Methods for Detection and Identification of Infectious Disease and Biological Warfare Agents

WISP-1 Binds to Decorin and Biglycan

Generating addressable protein microarrays with PROfusion™ covalent mRNA-protein fusion technology

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