Authentication Plans
On January 25, 2016, NIH implemented Rigor and Reproducibility requirements. As part of those requirements, applicants are required to provide a separate attachment, called "Authentication of Key Resources Plan". This plan is to outline the methods to ensure the identity and validity of key biological and/or chemical resources that will be used in the proposed studies.
Below is a list of resources that have been compiled from multiple resources and FAQs to help investigators develop their authentication plans.
The ABCs of Authentication Plans (NIH)
Learn more about authentication plans:
- What are “key biological and/or chemical resources”?
- How should authentication of key biological and/or chemical resources be addressed in grant applications?
- Example Authentication Plans
Things to know:
What is a key resource?
- May differ from laboratory to laboratory and/or over time
- May have qualities or qualification that could influence results
- Integral to the proposed research
- Includes resources no generated by NIH funds
What is not a key resource?
- Standard laboratory reagents that are not expected to vary over time
- Buffers
- Common chemical or biological reagents
- Datasets/databases
- Machinary
- Electronics
What are some examples of a key resource?
- Specialty chemicals
- Cell lines
- Antibodies
- Other biologics (especially for long-term storage)
Authentication Plan Resources
Antibodies are a frequently used tool but can vary batch-to-batch, including non-specific binding.1 Use a trusted manufacturer but antibodies should still be authenticated before experimentation and an authentication plan is still required for NIH grant submission.
Checklists and Validation Methods
Other Resources and Best Practices
- Improving Reproducibility: Best Practices for Antibodies from Sigma Aldrich
- Antybuddy.com is an independent antibody and protein review platform with supporting peer-to-peer Q&A forum
Literature Review
- Antibody Validation" by Jennifer Bordeaux, et. al.
- Standardize Antibodies used in Research by Andrew Bradbury and Andreas Plückthun
- A Proposal for Validation of Antibodies by Mathias Uhlen et. al.
1) Reproducibility crisis: Blame it on the antibodies by Monya Baker. Nature 2015; 521(7552):274-76
NIH suggest "applicants proposing to use cell lines could describe the method they plan to use to verify the identity and purity of the lines, which might include short tandem repeat (STR) profiling and mycoplasma testing." Below are some resources for writing an authentication plan and procedures for authenticating cell lines.
Cell lines should be obtained from a trusted vendor and a fresh cell line should be used before starting a series of experiments. Even using a trusted vendor, authentication plan is still required in NIH grant submission.
Trusted Vendors and Contaminated Cell Lines
Guidelines and Resources from Professional and Non-Profit Organizations
- Global Biological Standards Institute (GBSI)
- International Cell Line Authentication Committee (ICLAC)
Literature Review
- Know Thy Cells: Improving Biomedical Research Reproducibility by Leonard P. Freedman
- Changing the Culture of Cell Culture: Applying Best Practices and Authentication to Ensure Scientific Reproducibility by Leonard P. Freeman, Mark C. Gibson and Richard M. Neve
- Cell Line Authentication Demystified by Vivien Marx
- Standards for Cell Line Authentication and Beyond Jamie L. Almeida, Kenneth D. Cole and Anne L. Plant
- Recommendation of Short Tandem Repeat Profiling for Authenticating Human Cell Lines, Stem Cells, and Tissues by Rita Barallon, et. al.
- A Resource for Cell Line Authentication, Annotation and Quality Control by Mamie Yu, et. al.
Specific for Non-Human Cell Lines
- Authentication for non-human cell lines from ICLAC
- Mouse Cell Line Authentication by Jamie L. Almeida, Carolyn R. Hill and Kenneth D. Cole
Chemical analytical methods such as:
- Liquid or gas chromatography
- Mass spectrometry
- NMR
- Infrared spectroscopy
- Raman spectroscopy
- X-ray diffraction
- Etc.
https://grants.nih.gov/grants/guide/notice-files/NOT-OD-17-068.html
Genetically modified animals or cells might include PCR amplification or Southern blot to confirm genome modification.1
PCR Resources
- Learn more about polymerase chain reaction
- Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies by Todd C. Lorenz
Southern Blot Resources
- Southern Blotting by Ed Southern
1) https://grants.nih.gov/grants/guide/notice-files/NOT-OD-17-068.html
Authentication Plan FAQs
–It is expected to include a plan to independently verify the identity and activity of product before use
–If product is used long-term, consider the stability of the product and how validity of the product will be assessed over time
–If they’re using a “key resource,” may request information of authentication and include within own authentication plan
–Research conducted for resource development, including plans for validating the resource, should be described in Research Strategy section
–If proposing to collect primary cells for short-term culture as part of research, the activities (including plans for authentication identity of cells) should be described in Research Strategy
–If obtained from another laboratory, an authentication plan should be provided
–One-time analysis/sample? Do not need authentication plan
–Storing samples for repeated use/using stored samples? Authentication plan needed
–Using a “key resource” as part of imaging process, such as a proprietary dye? Authentication plan is needed
–Otherwise, the parameters to ensure reproducibility of imaging needs to be addressed as part of rigorous experimental design in Research Strategy
Yes!
Michelle Benson, PhD is available to answer your questions on authentication plans.
See also:
NIH & AHRQ: Rigor and Reproducibility Presentation by Michelle C. Benson and Stephanie F. Scott
NIH has provided some advice and examples for what may be included in an Authentication Plan, however there are currently no templates. Below are some example templates that investigators may find informative when developing their own plans.
If you are a Columbia investigator and would like to submit your Authentication Plan from a funded NIH award to Columbia's Authentication Plan Repository (login required), please email Michelle Benson at [email protected].
