Strategies to develop highly drug-tolerant cell-based neutralizing antibody assay: neutralizing antidrug antibodies extraction and drug depletion

Bioanalysis ◽  
2020 ◽  
Vol 12 (18) ◽  
pp. 1279-1293
Author(s):  
Zhihua Jiang ◽  
John Kamerud ◽  
Minlei Zhang ◽  
Carlos Cabrera Ruiz ◽  
Crisanto Guadiz ◽  
...  
Keyword(s):  
Sample Pretreatment ◽  
Neutralizing Antibody ◽  
Drug Tolerance ◽  
Acid Dissociation ◽  
Drug Interference ◽  
Drug Extraction ◽  
Alternative Approach ◽  
Method Optimization ◽  
Antidrug Antibodies ◽  
Novel Drug

Aim: Drug interference poses great analytical challenges for cell-based neutralizing antidrug antibodies (NAb) assay. The work aimed to improve assay drug tolerance through biotin-drug extraction with acid dissociation method optimization and developing new approach. Results: The NAb extraction with biotin-drug extraction with acid dissociation approach has been optimized by reducing biotinylated drug leaching and improving NAb elution efficiency, resulting in drug tolerance of up to 160 μg/ml. To circumvent the low acid elution efficiency of NAb from drug, a novel drug depletion approach was developed, which combined acid dissociation and drug targeted crosslinked capture, achieved drug tolerance up to 400 μg/ml. At last, a strategy workflow for sample pretreatment approach selection and optimization was established for improving drug tolerance of NAb assay. Conclusion: We demonstrated that reduced biotinylated drug leaching and the high NAb elution efficiency was critical for improving assay drug tolerance. Drug depletion offers an alternative approach to overcome low NAb elution efficiency.

scholarly journals Evaluation of Multiple Immunoassay Technology Platforms to Select the Anti-Drug Antibody Assay Exhibiting the Most Appropriate Drug and Target Tolerance

2016 ◽  
Vol 2016 ◽  
pp. 1-15 ◽  
Author(s):  
Justine Collet-Brose ◽  
Pierre-Jean Couble ◽  
Maureen R. Deehan ◽  
Robert J. Nelson ◽  
Walter G. Ferlin ◽  
...  
Keyword(s):  
Early Stage ◽  
Sample Pretreatment ◽  
Drug Tolerance ◽  
Development Stage ◽  
Appropriate Technology ◽  
Assay Development ◽  
Acid Dissociation ◽  
Technology Platform ◽  
Technology Platforms ◽  
Elisa Format

The aim of this study was, at the assay development stage and thus with an appropriate degree of rigor, to select the most appropriate technology platform and sample pretreatment procedure for a clinical ADA assay. Thus, ELISA, MSD, Gyrolab, and AlphaLISA immunoassay platforms were evaluated in association with target depletion and acid dissociation sample pretreatment steps. An acid dissociation step successfully improved the drug tolerance for all 4 technology platforms and the required drug tolerance was achieved with the Gyrolab and MSD platforms. The target tolerance was shown to be better for the ELISA format, where an acid dissociation treatment step alone was sufficient to achieve the desired target tolerance. However, inclusion of a target depletion step in conjunction with the acid treatment raised the target tolerance to the desired level for all of the technologies. A higher sensitivity was observed for the MSD and Gyrolab assays and the ELISA, MSD, and Gyrolab all displayed acceptable interdonor variability. This study highlights the usefulness of evaluating the performance of different assay platforms at an early stage in the assay development process to aid in the selection of the best fit-for-purpose technology platform and sample pretreatment steps.

Anti-Drug Antibody Responses to Lumiliximab Are Not Detected in Relapsed Refractory CLL Patients Treated with Lumiliximab in Combination with FCR in a Phase 1/II Study

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4210-4210 ◽  
Author(s):  
Julie Ranuio ◽  
Annalee Estrellado ◽  
Sarah Harris ◽  
Megan Shannon ◽  
Meena Subramanyam ◽  
...  
Keyword(s):  
Phase I ◽  
Colorimetric Detection ◽  
Drug Tolerance ◽  
Lymphocytic Leukemia ◽  
Acid Dissociation ◽  
Serum Samples ◽  
Drug Interference ◽  
Time Points ◽  
Coated Plate ◽  
Human Sera

Abstract Lumiliximab, an anti-CD23 monoclonal antibody that binds specifically to human CD23, is under investigation for the treatment of patients with relapsed refractory chronic lymphocytic leukemia (CLL). The clinical safety monitoring for Lumiliximab includes testing for the presence of anti-drug antibodies (ADAs) in patient serum samples. In order to determine if patients develop an ADA response to Lumiliximab, serum samples were collected at specific time points throughout a Phase I/II study (152–30) of Lumiliximab in combination with FCR in relapsed CLL. Patient sera from this Phase I/II study were tested in a standard bridging ELISA in which samples were added to a drug coated plate and ADAs were detected using conjugated drug and colorimetric detection. Validation results showed the sensitivity of the assay was 1000ng/ml using a purified polyclonal rhesus anti-Lumiliximab antibody in undiluted human sera. ADAs were not detected in any patients that had evaluable post treatment (Week 13, Week 25, Week 41, and Month 12 post first dose) serum samples in the 152–30 study. Over the last few years, there has been an effort in the immunogenicity field to develop ADA assays with increased sensitivity and to adapt formats that, in general, have increased drug tolerance, i.e. reduction of drug interference in the assay. Therefore, a new ADA assay was developed using Meso Scale Discovery (MSD) technology that has become the reliable and current method for detection of ADA responses and incorporates an acid dissociation step that has been shown in literature to improve drug tolerance. Using the new method the same evaluable samples from the phase I/II study were tested to determine if any previously unseen ADA responses could be detected. The samples were incubated with biotinylated and ruthenylated drug, added to a streptavidin coated plate and ADAs were detected using electrochemiluminescence. Validation results show that the newly developed MSD assay has sensitivity of 250ng/ml using a purified polyclonal chicken anti-Lumiliximab antibody in undiluted human sera. All the samples tested in the MSD assay (n=32) were negative despite greater sensitivity than the ELISA method and an acceptable drug tolerance at the time points samples were drawn. PK concentrations at the specified ADA collection times ranged between 0μg/ml and 114μg/ml whereas drug tolerance ranges from 1μg/ml to 200μg/ml depending on ADA concentrations. Drug interference is a major factor in not being able to detect antidrug responses, yet several of the corresponding post dose PK time points showed low to no drug levels and yet no anti-drug antibodies were detected. These data imply that Lumiliximab when administered in combination with FCR has a low immunogenicity rate.

scholarly journals High ionic strength dissociation assay (HISDA) for high drug tolerant immunogenicity testing

Bioanalysis ◽  
2020 ◽  
Author(s):  
Gregor Jordan ◽  
Alexander Pöhler ◽  
Florence Guilhot ◽  
Meike Zaspel ◽  
Roland F Staack
Keyword(s):  
Ionic Strength ◽  
Acid Treatment ◽  
Structural Integrity ◽  
Drug Tolerance ◽  
High Ionic Strength ◽  
Acid Dissociation ◽  
High Drug ◽  
Overnight Incubation ◽  
Antidrug Antibody ◽  
High Doses

Aim: Antidrug antibody (ADA) assessment may be challenged in studies that involve the administration of high doses of biotherapeutics and/or with long half-lives. In such cases, ADA assays with optimized drug tolerance are desired. Material & Methods: We evaluated the use of MgCl2 to develop high ionic strength dissociation assays in two investigational examples (bridging enzyme-linked immunosorbent ADA assays) to attain high drug tolerance while maintaining best possible structural integrity of ADAs. Results: Both ADA-bridging assays treated with MgCl2 showed improved drug tolerance and higher signal-to-blank values compared with overnight incubation or acid treatment. Conclusion: The use of MgCl2 treatment in ADA-bridging assays provides a sensitive, drug tolerant and easy-to-use alternative in cases where acid dissociation is not possible or unwanted.

scholarly journals Drug Repositioning: New Approaches and Future Prospects for Life-Debilitating Diseases and the COVID-19 Pandemic Outbreak

Viruses ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 1058
Author(s):  
Zheng Yao Low ◽  
Isra Ahmad Farouk ◽  
Sunil Kumar Lal
Keyword(s):  
Drug Development ◽  
De Novo ◽  
Drug Repositioning ◽  
Personalised Medicine ◽  
Technological Advancement ◽  
The Novel ◽  
Ongoing Research ◽  
Alternative Approach ◽  
Good Potential ◽  
Novel Drug

Traditionally, drug discovery utilises a de novo design approach, which requires high cost and many years of drug development before it reaches the market. Novel drug development does not always account for orphan diseases, which have low demand and hence low-profit margins for drug developers. Recently, drug repositioning has gained recognition as an alternative approach that explores new avenues for pre-existing commercially approved or rejected drugs to treat diseases aside from the intended ones. Drug repositioning results in lower overall developmental expenses and risk assessments, as the efficacy and safety of the original drug have already been well accessed and approved by regulatory authorities. The greatest advantage of drug repositioning is that it breathes new life into the novel, rare, orphan, and resistant diseases, such as Cushing’s syndrome, HIV infection, and pandemic outbreaks such as COVID-19. Repositioning existing drugs such as Hydroxychloroquine, Remdesivir, Ivermectin and Baricitinib shows good potential for COVID-19 treatment. This can crucially aid in resolving outbreaks in urgent times of need. This review discusses the past success in drug repositioning, the current technological advancement in the field, drug repositioning for personalised medicine and the ongoing research on newly emerging drugs under consideration for the COVID-19 treatment.

Spectroscopic Determination of Acid Dissociation Constants of Some Novel Drug Precursor 6-Acylbenzothiazolon Derivatives

2010 ◽  
Vol 55 (11) ◽  
pp. 4752-4756 ◽  
Author(s):  
Yadigar Gülseven Sıdır ◽  
İsa Sıdır ◽  
Halil Berber ◽  
Erol Taşal ◽  
Cemil Öğretir
Keyword(s):  
Dissociation Constants ◽  
Acid Dissociation ◽  
Spectroscopic Determination ◽  
Acid Dissociation Constants ◽  
Novel Drug

A bridging immunogenicity assay for anti-cabiralizumab antibodies: overcoming the low assay cut point and drug tolerance challenges

Bioanalysis ◽  
2021 ◽  
Author(s):  
Long Yuan ◽  
Carol R Gleason ◽  
Dennis Stocker ◽  
Li Li ◽  
Jim X Shen ◽  
...  
Keyword(s):  
Clinical Studies ◽  
Drug Tolerance ◽  
Normal Serum ◽  
Acid Dissociation ◽  
Reagent Concentration ◽  
Acid Pretreatment ◽  
Cut Points ◽  
Cut Point ◽  
Immunogenicity Assay ◽  
Assay Conditions

Background: To support the clinical studies of cabiralizumab, an immunogenicity assay for detecting anti-cabiralizumab antibodies is required. Results: Strategies were developed to overcome two major bioanalytical challenges: poor drug tolerance of the anti-drug antibodies assay and very low cut point observed in the screening and confirmatory assays. By using acid dissociation (400 mM glycine solution at pH 2.0), drug tolerance of 200 μg/ml drug was achieved for both the screening and confirmatory assays. Effects of biological matrix (disease state vs normal serum) and assay conditions (capture/detector reagent concentration, minimum required dilution, acid pretreatment) on assay cut points were systematically evaluated. Conclusion: A bridging immunogenicity assay for detecting anti-cabiralizumab antibodies in human serum has been successfully developed, validated and applied to clinical studies.

Nanomedicine: A Promising Avenue for the Development of Effective Therapy for Breast Cancer

2020 ◽  
Vol 20 (8) ◽  
pp. 603-615
Author(s):  
Ali Sartaj ◽  
Sanjula Baboota ◽  
Javed Ali
Keyword(s):  
Breast Cancer ◽  
Side Effects ◽  
Relevant Literature ◽  
High Dose ◽  
Surgical Interventions ◽  
Alternative Approach ◽  
Novel Drug ◽  
Low Solubility ◽  
Pass Metabolism ◽  
Promising Avenue

Purpose: Breast cancer is the most probable cancer among women. However, the available treatment is based on targeting different stages of breast cancer viz., radiation therapy, hormonal therapy, chemotherapy, and surgical interventions, which have some limitations. The available chemotherapeutics are associated with problems like low solubility, low permeability, high first-pass metabolism, and P-glycoprotein efflux. Hence, the aforementioned restrictions lead to ineffective treatment. Multiple chemotherapeutics can also cause resistance in tumors. So, the purpose is to develop an effective therapeutic regimen for the treatment of breast cancer by applying a nanomedicinal approach. Methods: This review has been conducted on a systematic search strategy, based on relevant literature available on Pub Med, MedlinePlus, Google Scholar, and Sciencedirect up to November 2019 using keywords present in abstract and title of the review. As per our inclusion and exclusion criteria, 226 articles were screened. Among 226, a total of 40 articles were selected for this review. Results: The significant findings with the currently available treatment is that the drug, besides its distribution to the target-specific site, also distributes to healthy cells, which results in severe side effects. Moreover, the drug is less bioavailable at the site of action; therefore, to overcome this, a high dose is required, which again causes side effects and lower the benefits. Nanomedicinal approaches give an alternative approach to avoid the associated problems of available chemotherapeutics treatment of breast cancer. Conclusion: The nanomedicinal strategies are useful over the conventional treatment of breast cancer and deliver a target-specific drug-using different novel drug delivery approaches.

scholarly journals Novel methodology comparing two assay formats for the assessment of immunogenicity from clinical trial samples

Bioanalysis ◽  
2021 ◽  
Vol 13 (14) ◽  
pp. 1113-1121
Author(s):  
Daniel A Peterson ◽  
Thomas G Pottanat ◽  
Heather Denning ◽  
Nicoletta Bivi ◽  
John H Sloan ◽  
...  
Keyword(s):  
Clinical Trial ◽  
Clinical Trials ◽  
Human Serum ◽  
Drug Tolerance ◽  
Interlaboratory Comparisons ◽  
Clinical Interpretation ◽  
Blinded Fashion ◽  
Assay Format ◽  
Antidrug Antibodies ◽  
Immunogenicity Assays

Aim: We present a novel methodology to compare results between distinct immunogenicity assays, performed by two laboratories, for the same biotherapeutic. Materials & methods: Human serum pools from clinical trials were generated to provide representative immunogenicity titers. Pools were evaluated at two laboratories in a blinded fashion to assess the effect of assay format and laboratory change on clinical interpretation of immunogenicity results. Results: The laboratories validated two different assay formats and demonstrated comparable sensitivity and drug tolerance. Overall, the comparisons in assay format and laboratory ensured a comparable ability to detect treatment-emergent antidrug antibodies for a biotherapeutic. Conclusion: We have established an approach, using pooling of patient samples, that allows for the interlaboratory comparisons without creating duplicative results.

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