Multiplex autoantibody profiling for autoimmune diseases and cancer
This presentation is the property of its rightful owner.
Sponsored Links
1 / 25

Multiplex autoantibody profiling for autoimmune diseases and cancer PowerPoint PPT Presentation


  • 117 Views
  • Uploaded on
  • Presentation posted in: General

Multiplex autoantibody profiling for autoimmune diseases and cancer. April, 2, 2014. Profile antibodies to over 10,000 human proteins using Luminex xMAP™ technology. Jim Lazar, Ph.D. VP, Assay Development. OriGene Introduction. OTI Locations Rockville (HQTs/R&D) Seattle (Blue Heron)

Download Presentation

Multiplex autoantibody profiling for autoimmune diseases and cancer

An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -

Presentation Transcript


Multiplex autoantibody profiling for autoimmune diseases and cancer

Multiplex autoantibody profiling for autoimmune diseases and cancer

April, 2, 2014

Profile antibodies to over 10,000 human proteins using Luminex xMAP™ technology

Jim Lazar, Ph.D.

VP, Assay Development


Origene introduction

OriGene Introduction

  • OTI Locations

    • Rockville (HQTs/R&D)

    • Seattle (Blue Heron)

    • Delaware (SDIX)

    • Maine (SDIX)

    • Foster City (BioCheck)

    • Beijing

    • Wuxi


Antibodies as biomarkers

Antibodies as Biomarkers

  • The immune system functions as a an early warning system.

    • Malignant transformation, infections, and many diseases can trigger an immune response

  • Antibodies are abundant, easily detectable and stable in serum.


Autoantibodies to tumor antigens

Autoantibodies to Tumor Antigens

  • Tumors typically over-express multiple normal and mutant antigens

  • Autoantibodies to tumor antigens can be detected much earlier than other cancer biomarkers

  • Frequency of autoantibody responses to an individual tumor antigen is typically 15-20%.

  • Responses to a panel of antigens can be higher than 90%

  • Multiplexed autoantibody detection to a panel of antigens is necessary.

What is EarlyCDT–Lung?

EarlyCDT-Lung is a simple, physician-ordered, diagnostic blood test that measures the presence of autoantibodies to a panel of seven lung cancer-associated antigens.


Autoimmune diseases

Autoimmune Diseases

  • Autoimmune diseases affect an estimated 3% of the world population.

  • Over 150 recognized autoimmune diseases.

  • Each disease is characterized by autoantibodies to multiple antigens.

  • Profiles may help classify disease variants and indicate organ-specific involvement.

  • Autoantibody profiles may help guide the development of antigen-specific tolerizing therapy.


Biomarker discovery

Biomarker Discovery

  • Proteomic /2D gel methods

  • Proteomic Mass Spec methods

  • High Density Protein Microarrays

Identification of many targets from a small number of samples

Biomarker Screening

& Quantitative Evaluation

Large sample #s

Refine target list

ROC calculations

  • Luminex Bead Arrays

  • Mid-Density Multiplexed methods

Clinical Assays

  • Luminex Bead Arrays

  • Low-Density Multiplexed methods


10 000 full length human proteins

10,000+ Full Length Human Proteins

  • Produced from TrueORF Gold™ cDNA clones

    • Fully sequenced

    • Expression validated

  • Expressed in HEK293 cells

    • Human-expressed

    • Affinity purified

  • Native presentation

    • Optimal preservation of protein structure,

    • post-translational modifications


Available antigens

Available Antigens

Any of OriGene’s >10,000 purified proteins can be custom-coupled to Luminex beads


Luminex xmap technology

Luminex xMAP™ technology

  • Multiplex Bead Array

  • Efficient high-throughput microplate format

  • Up to 100 targets per well

  • OriGene is a Luminex partner and Certified Assay Developer


Assay principle

Assay Principle

+

Protein-coupled Luminex Beads

Sample containing human IgG

+

Mix with Anti-human PE Conjugate

Read Signal in Luminex


Product configuration

Product Configuration

  • TruePlex™ Human Antibody Profiling Kit

    • Assay Reagents, Control Bead Mix, Positive Control

combine with

  • Custom TruePlex™ Antibody Profiling Array

  • TruePlex™ Antibody Profiling Array

  • (pre-defined array)

and/or

Multiple profiling arrays can be mixed together and analyzed as a larger multiplex


Assay protocol

Assay Protocol

Dilute sample

Mix with beads, incubate

Total time – about 2.5 hours for 96 samples

Up to 100 results per well

Quantitative results

Wash, add conj. & incubate

Wash

Read in Luminex

Analyze Data


Stringency options

Stringency Options

  • Sample Diluent can be prepared at different stringencies

    • High, medium or low

  • High-stringency Sample Diluent will minimize non-specific signals and is recommended for initial sample testing.

  • The medium- or low-stringency Sample Diluent may give better results for samples

    • with low autoantibody levels,

    • in which the antibody affinity may be low

    • for higher sample dilutions (1:500 or greater),


Available trueplex profiling arrays

Available TruePlex™ Profiling Arrays


Autoimmune profiles in serum

Autoimmune profiles in serum


Autoimmune sera data from ssb and trim21

Autoimmune sera – data from SSB and TRIM21


Normal non disease sera

Normal (non-disease) Sera


Dynamic range

Dynamic Range


Control results

Control Results


Breast cancer samples tested with 20 plex tumor antigen panel

Breast cancer samples tested with 20-plex tumor antigen panel


Available cancer tumor antigens known to generate autoantibodies

Available cancer tumor antigens known to generate autoantibodies

  • Breast Cancer

    • ASB-9, BAT4, BDNF, Survivin, Livin, BMX, BRCA2, P16, CSNK1E, NY-ESO-1/LAGE1, CTBP1, DBT, EIF3E, HER2, Fibulin, FKBP4, GIPC-1, HSP-90, HSPA4, HSP-27, HSP-60, IMP1 , P62, IMP2, MUC1, C-Myb, c-myc, PDCD6IP, PPIA, PRDX2, RAB5A, RAC3, Lipophilin B, SERPINH1, SF3A1, SOX2, p53, TRIM32, UBAP1

  • ProstateCancer

    • Caldesmon 1, Clusterin, c-myc, HER2/neu, HSP70, HSP71, IMP1, MCP1, NY-ESO-1/LAGE1, p53, p62, p90, PARK7, PSA, RACE, SSX2, SSX4, Survivin, TARDBP, TTLL12

  • Pancreatic Cancer

    • CTDSP1, DNAJB1,EIF4A3 (DDX48), ELAC1,GAS2, HCFC1R1,HERPUD1, MAPK9, NR2E3, NR2E3, PDLIM1 (CLP36), PGK1,PPARG, PTPRA, RNF213 (KIAA1618), ROR2, SHOC2, SMOX, TMOD1, TMSB10, ULK4

  • Antigens for other cancers and diseases available including

    • Ovarian cancer, melanoma, hepatocellular carcinoma, lung cancer, diabetes, COPD, multiple sclerosis

http://www.origene.com/Luminex/Proteinarray


Extensive bibliography available

Extensive Bibliography Available

Autoantibodies to tailor-made panels of tumor-associated antigens in breast carcinoma.E Piura and B PiuraJ Oncol, Jan 2011; 2011: 982425.

Seromic profiling of ovarian and pancreatic cancer.

S. Gnjatic S, et al.

PNAS U S A. Mar 2010 16;107(11):5088-93.

A Distinct Repertoire of Autoantibodies in Hepatocellular Carcinoma Identified by Proteomic Analysis

François Le Naour, et al.

Mol. Cell. Proteomics, Mar 2002; 1: 197 - 203.

Multiple serological biomarkers for colorectal cancer detection.

CC Chan, et al.

Int J Cancer, Apr 2010; 126(7): 1683-90.


Summary

Summary

  • Autoantibodies are useful biomarkers for cancer and autoimmune diseases.

  • OriGene’s TruePlex profiling kits are optimally suited for biomarker screening and validation.

  • Choose from over 10,000 proteins expressed in human cells.

  • The assay procedure is simple and fast.

  • Make great discoveries

  • Improve human health


Thank you for attending

Thank you for attending !

Questions ?

Contact us:

[email protected]

[email protected]

240-620-0237 (direct)

www.origene.com/assays


  • Login