Layered Peptide Arrays

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Layered Peptide Arrays Gallya Gannot, Michael A. Tangrea, John W. Gillespie, Heidi S. Erickson, Benjamin S. Wallis, Rose Anne Leakan, Vladimir Knezevic, Dan P. Hartmann, Rodrigo F. Chuaqui, Michael R. Emmert-Buck The Journal of Molecular Diagnostics Volume 7, Issue 4, Pages 427-436 (October 2005) DOI: 10.1016/S1525-1578(10)60573-9 Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Schematic of LPA system. Each membrane is coated with a different peptide or antigen containing an epitope for the antibody of interest. Antibodies move from the multiwell grid and through the membranes where they are captured by their corresponding antigens. Labeled secondary antibodies are used for detection. The Journal of Molecular Diagnostics 2005 7, 427-436DOI: (10.1016/S1525-1578(10)60573-9) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 LPA prototype 1: antibody control experiment. Demonstration of successful antibody capture using a 10-layer LPA system. The Journal of Molecular Diagnostics 2005 7, 427-436DOI: (10.1016/S1525-1578(10)60573-9) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 LPA prototype 1: serum samples. A: Schematic drawing of LPA system. Each color designates a different peptide coating. B: Bar graph showing LPA analysis of patient samples and controls. The Journal of Molecular Diagnostics 2005 7, 427-436DOI: (10.1016/S1525-1578(10)60573-9) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 4 Comparison of LPA prototype 1 with standard ELISA. A: Comparison of SSA antibody titer. B: Comparison of SSB antibody titer. The Journal of Molecular Diagnostics 2005 7, 427-436DOI: (10.1016/S1525-1578(10)60573-9) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 5 LPA prototype 2: antibody control experiment. A: Schematic drawing of the system. B: Table showing organization of samples, and images of membranes showing successful capture of antibodies. C: CAR antibody capture on membranes 6, 16, 26, 36, and 46 (from top to bottom). D: Quantitative measurement of signal intensity on each membrane showing twofold dilution of samples. The Journal of Molecular Diagnostics 2005 7, 427-436DOI: (10.1016/S1525-1578(10)60573-9) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 6 LPA prototype 2: serum samples. Comparison with standard ELISA. A: Table showing comparison of the LPA system and a standard ELISA for SSB antibody titer. Thirty-two Sjögren's syndrome patients and eight healthy volunteers were analyzed. B: Quantitative measurements of SSB antibody analysis using an ELISA or the LPA system. The Journal of Molecular Diagnostics 2005 7, 427-436DOI: (10.1016/S1525-1578(10)60573-9) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 7 Specific antibody detection in sera and saliva. A: Measurement of antibodies in sera and saliva for SS patients (P) or controls (NV). B: Analysis of variance values for SS patients and normal volunteers in serum and saliva samples. The Journal of Molecular Diagnostics 2005 7, 427-436DOI: (10.1016/S1525-1578(10)60573-9) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 8 PCA clustering for patients (P) and controls (N). PCA mapping of serum and saliva antibodies for 32 SS patients (P) and 8 controls (N). The Journal of Molecular Diagnostics 2005 7, 427-436DOI: (10.1016/S1525-1578(10)60573-9) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions