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  • 1.  Strategies to mitigate ultra-low cut points

    Posted 10-01-2025 22:13

    Hello everyone,

    I would appreciate hearing your perspectives on strategies to avoid ultra-low ADA screening cut points. I have encountered discussions suggesting the use of a minimum cut point (for example, a cut point of 1.2 to incorporate the accepted CV allowance), even if that causes the FPR to fall below 5%, and leads to missing some low-titer, inconsequential ADA responses. Can anyone share their experiences/strategies on this topic?

    Additionally, are there any publications/guidances on this topic that I may have missed?

    Thank you

    Arkadeep



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    Arkadeep Sinha, PhD
    Director, Bioanalytical Sciences
    Upstream Bio
    [email protected]

    Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.
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  • 2.  RE: Strategies to mitigate ultra-low cut points

    Posted 10-02-2025 12:04

    Please include me in the discussion. [email protected]



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    Bin Xu Ph.D.
    CEO
    ABX Biotech LLC
    Ambler PA
    [email protected]

    Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.
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  • 3.  RE: Strategies to mitigate ultra-low cut points

    Posted 10-02-2025 15:47

    Hi Arkadeep,

    Ultra-low ADA screening cut points are indeed a critical area in immunogenicity assessments. I believe there are several perspectives that can help address the challenges associated with overly sensitive thresholds.
    First, to ensure ADA assays are both clinically relevant and scientifically robust, it's essential to start with the method development strategy. Assays should be designed to balance both sensitivity and specificity, being able to detect "real" ADA positive responses while minimizing false positives due to assay background or biological variability. 
    Second, alternative cut point strategies, such as the minimum 1.2, are a valid approach and can help avoid the ultra-low cut points. However, this may require adjusting the false positive rate threshold to something lower, such as 1%. Additionally, not removing biological outliers during the cut point determination could help raise the background levels while also better incorporating the potential heterogeneity of the patient population. 
    Lastly, there are alternative ADA interpretation approaches that go beyond the classical binary cut point determination. Evaluating ADA responses in the context of the entire patient profile allows for a more nuanced understanding of the potential clinical impacts. Using signal-to-noise (S/N) ratios as a continuous variable, similar to biomarker assays, avoids the need for a fixed cut point and enables integration of ADA data with other datasets to assess clinical impact more holistically. 
    Any approach to ADA assay design and interpretation should be grounded in the immunogenicity risk assessment of the molecule. This ensures that the monitoring strategy is tailored to identify clinically relevant ADA responses. At Immunologix, this is a frequent topic of discussion among our team and with our drug development partners. We've found that each program presents unique challenges, and that open, collaborative dialogue is key to developing a phase-appropriate and risk-based strategy for both immunogenicity monitoring and assay design. These conversations often lead us to the most effective and scientifically sound solutions.
    Thanks
    RJ


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    Robert Neely Ph.D.
    Scientific Director-Translational Sciences
    Immunologix Laboratories
    Mount Laurel NJ
    [email protected]

    Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.
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  • 4.  RE: Strategies to mitigate ultra-low cut points

    Posted 10-04-2025 10:16

    Hello Arkdeep,

    I'm posting this on the Immunogenicity board and the Bioanalytical board to cover both communication streams.  Apologies to those of you that are reading this twice.  John Kamerud and Robert Neely all share excellent points regarding ultra-low cut points.  You should definitely check to make sure you aren't excluding too many outliers (I start paying attention above 10% and get nervous if >15% are excluded), and making sure your negative control is representative of the cut point individuals.  

    The timing of your question is excellent as I'm working on a collaborative paper with two CROs and BioData Solutions to discuss this topic in greater detail.  Our conclusion is that ultra-low cut points are not bad at all.  They don't lead to inflated rates of immunogenicity and runs don't fail more often than those with higher cut points.  Outside the scope of the manuscript's discussion is the clinical interpretation of immunogenicity and Robert Neely brings up a good point that reporting S/N can add nuanced information about the immune response superior to Positive/Negative/Titer. 

    Here is a sneak peak of what is in the paper:  We tabulated cut points from 185 assays and over half of the assays had cut points at 1.20 or lower and a third of assays had cut points less than or equal to 1.10, thus ultra-low.  We performed in-depth analysis of ~12 ultra-low cut point case studies using various assay formats and drug modalities and only two required in-study cut points. In both cases there were assignable causes for the ultra-low cut points, rather than low assay and biological variability.  The first case was overzealous removal of outliers (>15%) and the second case was a negative control that was above the cut point individuals' responses.  We presented a portion of this data at EBF, so if you'd like a copy of the poster, please let me know.  

    In short, ultra-low cut points were found to be quite robust and shouldn't cause undue concern for the bioanalytical scientists as long as an excessive number of outliers are not excluded, and the negative control is representative of the population.   

    Robert



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    Robert Kernstock
    Principal Scientist
    Astellas Research Institute of America LLC
    Northbrook IL
    [email protected]

    Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.
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  • 5.  RE: Strategies to mitigate ultra-low cut points

    Posted 10-06-2025 08:17

    Robert,

    Can you please share this poster with me? Thanks.

     We presented a portion of this data at EBF, so if you'd like a copy of the poster. 



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    [email protected]

    Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.Associate Scientific DirectorTakeda
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  • 6.  RE: Strategies to mitigate ultra-low cut points

    Community Leadership
    Posted 10-06-2025 09:59

    @Robert Kernstock, while your summation indicates lack of undue concern from the perspective of the bioanalytical scientist, the real concern that Arkadeep raises is from the perspective of the drug program and misleading perception of high rates of immunogenicity when the bulk of responses are clinically irrelevant.  This creates untold swirl within organizations and consumes time, money and resources to appropriately message to key stakeholders, investors and regulators.  The cost to industry is substantial with no added benefit to patients.  In every case where we have re-analyzed immunogenicity data sets from clinical studies using S/N, we have been able to provide more complete, clearer narratives that more quickly lead to identification of responses that have any clinical impact.  



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    Lauren Stevenson Ph.D.
    Chief Scientific Officer
    Immunologix Laboratories
    Tampa FL
    [email protected]

    Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.
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  • 7.  RE: Strategies to mitigate ultra-low cut points

    Posted 10-07-2025 08:04
    My thoughts exactly, Lauren. We are often reacting to the concerns of management who fear the word “immunogenicity” is the kiss of death to their program, when in fact many very successful products have reported a high incidence of ADA.

    John Kamerud
    JK Bioanalytical Consulting LLC

    Sent from my iPhone




  • 8.  RE: Strategies to mitigate ultra-low cut points

    Posted 10-04-2025 18:11

    Thank you, Robert, for your detailed explanation on this topic. To be fair, my thoughts on this subject were prompted largely by recent discussions initiated by folks at Immunologix. To clarify my question further, my concern is not about the assay's performance or its ability to appropriately capture population variability, as monitoring the FPR can answer that. Rather, my concern lies with the low-titer true positives that are detected as a result of these ultra-low cut points. In many cases, these true low-titer ADA positives appear to lack clinical relevance. Given the recent emphasis on identifying clinically meaningful ADA responses and the broader discussions around shifting the testing paradigm, my post was intended to question whether it might be time to re-evaluate the relevance of maintaining such ultra-low cut points.

    Your suggestion of using S/N is definitely one way of getting around this.

    Thank you

    Arkadeep



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    Arkadeep Sinha, PhD
    Director, Bioanalytical Sciences
    Upstream Bio
    [email protected]

    Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.
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  • 9.  RE: Strategies to mitigate ultra-low cut points

    Posted 10-07-2025 21:23

    Arkdeep, Lauren, John, Robert N., Joleeen, and Eric,

    Once again, I'm replying on both discussion boards. I'm so thankful for your initial post and all of the responses so far.  

    The publication on ultra-low cut points, as Lauren suggested, is targeted toward the Bioanalytical Scientist to give them confidence that their cut point factor is solid (reliable and assay appropriate) as long as they assess the outlier removal and NC response vs the population.  I couldn't agree more that the approach towards the assessment of unwanted immunogenicity could use a refresh.  I love the idea of looking at S/N, which in most cases follows Titer, as a means of teasing out impact of ADA on pharmacokinetics, safety, and efficacy.  Additionally, S/N has the advantage of a single sample measurement in the screening assay rather than going through the three-tiered approach cutting down on unnecessary sample analysis costs and timelines.  If I may be so bold, S/N is a surrogate for ADA concentrations, just like titer is, and who knows, in the future immunogenicity assays may have a standard curve applied to them resulting in relative ADA 'concentrations'.  To be clear, immunogenicity assays are not PK assays, they are biomarker assays, and many of the challenges with biomarker assays regarding reference standards would apply to quantitative immunogenicity assays.  I'm also not advocating for such an approach, but it is one that could be taken.  

    Now to address Arkdeep's original point about clinical impact. I would say that in the case studies we explored, the rates of immunogenicity were not elevated, but I can't say for certain if those positive immunogenicity results were clinically impactful.  My guess is they were not. I'd also say that immunogenicity assessments are almost always best understood with supplemental data in retrospect.  It would be nearly impossible to say that a cut point is appropriate before conducting the clinical study (see Joleen's comment).  In my situation, and probably similarly to what is being done with Lauren at Immunologix, you start stratifying the ADA responses with PK, efficacy, and safety data.  For example, does an ADA S/N < 3 (or titers < 400) impact drug concentrations?  If not, then you've got great supporting data to suggest that while 'low-level' ADA may be detected, below a certain threshold these ADA have no apparent clinical safety/efficacy impact. Then you can state things like 20% of subjects developed ADA, but only 5% developed ADA that led to decreased efficacy.  

    It is a shame that there is so much swirl when we are reporting high rates of immunogenicity where the bulk of the data shows that they are not impactful (summarizing Lauren's comment).  I've been on programs where immunogenicity results led to product termination when there were no data showing impact on efficacy or safety, nevertheless "rate of immunogenicity" was a marketing concern.  I'm so glad that Bioanalytical scientists (and AAPS) are having these discussions and publishing white papers and other examples demonstrating the use of alternative strategies to summarize the impact of ADA (e.g. S/N vs Titer, and utility of confirmatory assays).  I hope that smart science continues to lead us towards better outcomes.  



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    Robert Kernstock
    Principal Scientist
    Astellas Research Institute of America LLC
    Northbrook IL
    [email protected]

    Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.
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