1. How antibodies can prevent medical progress and how they can be great tools. Birmingham 2015

2. What do we expect?

3. What do we get?

4. Recent literature on antibody performance F1000Research F1000 recommended OPINION ARTICLE published 15 Oct 2014 Commercial antibodies and their validation [v2; ref status: indexed, http://f1000r.es/4jp]http://f1000r.es/4jp JLA Voskuil F1000Research OPINION ARTICLE published 28Apr 2015 How difficult is the validation of clinical biomarkers? [v1; ref status: indexed, http://f1000r.es/5al ]http://f1000r.es/5al Jan Voskuil

5. Recent attacks from Nature magazine NATURE | NEWS Irreproducible biology research costs put at $28 billion per year Study calculates cost of flawed biomedical research in the United States. Monya Baker 09 June 2015 NATURE | TOOLBOX Researchers argue for standard format to cite lab resources Research Resource Identifier (RRID) aims to clean up poorly referenced data. Dalmeet Singh Chawla 29 May 2015 NATURE | NEWS FEATURE Reproducibility crisis: Blame it on the antibodies Antibodies are the workhorses of biological experiments, but they are littering the field with false findings. A few evangelists are pushing for change. Monya Baker 19 May 2015 NATURE | COMMENT Reproducibility: Standardize antibodies used in research To save millions of dollars and dramatically improve reproducibility, protein-binding reagents must be defined by their sequences and produced as recombinant proteins, say Andrew Bradbury, Andreas Plückthun and 110 co-signatories. Andrew BradburyAndrew Bradbury & Andreas PlückthunAndreas Plückthun 04 February 2015

6. How antibodies prevent progress in biomedical science How antibodies can be used as powerful tools Alternatives to antibodies: Affimers/Affibodies Aptamers

7. Examples of Antibodies on the market appearing as different but they are not proteinBiocompare (suppliers)actual # different Ab AIF1 / ABI1348 (31)~30 PIC3CA220 (35)~32 FOXP31055 (47)~31 NANOG574 (42)~45 OCT4.582 (41)~30 PIK3CA#frequency Acris16 AOL10 LSBIO10 Abcam97./4 Abnova65./1 Novus64./3./2 Thermo6 Abgent59./5 Sigma5 Biorbyt5 GeneTex47./6 ProteinTech44./2 Aviva3 CST32./1./1 Mybiosource3 Millipore21 Origene21./1 SantaCruz21./1 EverestBiotech116 ABDSerotec1 ProSci1 AIF1/ABI1ACRIS24 products speciesantigenmanufrfrequency goataa81-9315 goataa70-8014 goataa28-4215 rabbitprotein21 rabbitaa13-4421 rabbitaa85-14731 rabbitpeptide31 rabbitpeptide41 mousemab#102252 mousemab#2A2-B651

8. Other External Manufacturer (Original Equipment Manufacturer) Welcome to the world of OEM One manufacturer, one product with many brands QC data responsibility of rebranding vendor Batch-to-batch variations ignored by many vendors QC data presented from previous animals QC data presented from own customers QC data presented from own QC department Rebranding vendors buying from each other Same product appearing several times on same catalogue

9. How antibodies prevent progress in biomedical science How antibodies can be used as powerful tools Alternatives to antibodies: Affimers/Affibodies Aptamers

10. Specificity Entire protein as antigen Specific region as antigen Peptide as antigen

11. Shared epitopes Cross reactivity of clinically validated anti-HER2 antibodies Schrohl A et al, Abstract for SABCS 2010, Abstract No. 850345 HER2 antibodies HER1HER2HER3HER4 Herceptest™ (Dako) v PATHWAY® (Vantana) vv Oracle™ (Leica) vv Conclusion Avoid antibodies with unknown epitopes