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In JoVE (1)
Other Publications (1)
Articles by Kelly Lamiman in JoVE
High Throughput Sequential ELISA for Validation of Biomarkers of Acute Graft-Versus-Host Disease
Bryan Fiema*1, Andrew C. Harris*1, Aurelie Gomez1, Praechompoo Pongtornpipat1, Kelly Lamiman1, Mark T. Vander Lugt1, Sophie Paczesny1
1Pediatric Blood and Marrow Transplant Program, University of Michigan
High throughput validation of multiple candidate biomarkers can be performed by sequential ELISA in order to minimize freeze/thaw cycles and use of precious plasma samples. Here, we demonstrate how to sequentially perform ELISAs for six different validated plasma biomarkers1-3 of graft-versus-host disease (GVHD)4 on the same plasma sample.
Published October 31, 2012. Keywords: Medicine, ELISA, Sequential ELISA, Cytokine, Blood plasma, biomarkers, proteomics, graft-versus-host disease, Small sample, Quantification
Other articles by Kelly Lamiman on PubMed
New Ligation-independent Cloning Vectors Compatible with a High-throughput Platform for Parallel Construct Expression Evaluation Using Baculovirus-infected Insect Cells
Protein Expression and Purification. May, 2011 | Pubmed ID: 21262364
Biomedical research has undergone a major shift in emphasis over the past decade from characterizing the genomes of organisms to characterizing their proteomes. The high-throughput approaches that were successfully applied to sequencing of genomes, such as miniaturization and automation, have been adapted for high-throughput cloning and protein production. High-throughput platforms allow for a multi-construct, multi-parallel approach to expression optimization and construct evaluation. We describe here a series of baculovirus transfer and expression vectors that contain ligation-independent cloning regions originally designed for use in high-throughput Escherichia coli expression evaluation. These new vectors allow for parallel cloning of the same gene construct into a variety of baculovirus or E. coli expression vectors. A high-throughput platform for construct expression evaluation in baculovirus-infected insect cells was developed to utilize these vectors. Data from baculovirus infection expression trials for multiple constructs of two target protein systems relevant to the study of human diseases are presented. The target proteins exhibit a wide variation in behavior and illustrate the benefit of investigating multiple cell types, fusion partners and secretion signals in optimization of constructs and conditions for eukaryotic protein production.