scholarly journals High-Throughput Epitope Binning Assays on Label-Free Array-Based Biosensors Can Yield Exquisite Epitope Discrimination That Facilitates the Selection of Monoclonal Antibodies with Functional Activity

PLoS ONE ◽  
2014 ◽  
Vol 9 (3) ◽  
pp. e92451 ◽  
Author(s):  
Yasmina Noubia Abdiche ◽  
Adam Miles ◽  
Josh Eckman ◽  
Davide Foletti ◽  
Thomas J. Van Blarcom ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
High Throughput ◽  
Functional Activity ◽  
Label Free ◽  
Selection Of

scholarly journals A novel approach for rapid high-throughput selection of recombinant functional rat monoclonal antibodies

2018 ◽  
Vol 19 (1) ◽  
Author(s):  
Qin Chen ◽  
Shengping Qiu ◽  
Huanhuan Li ◽  
Chaolong Lin ◽  
Yong Luo ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
High Throughput ◽  
Novel Approach ◽  
Selection Of

Characterization of two in vivo challenge models to measure functional activity of monoclonal antibodies to Plasmodium falciparum circumsporozoite protein

2020 ◽  
Author(s):  
Rama Raghunandan ◽  
Bryan T Mayer ◽  
Yevel Flores-Garcia ◽  
Monica W Gerber ◽  
Raphael Gottardo ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Monoclonal Antibodies ◽  
Functional Activity ◽  
Statistical Power ◽  
Rank Order ◽  
Circumsporozoite Protein ◽  
Dose Selection ◽  
Selection Of

Abstract Background New strategies are needed to reduce the incidence of malaria, and promising approaches include the development of vaccines and monoclonal antibodies (mAbs) that target the circumsporozoite protein (CSP). To select the best candidates and speed development, it is essential to standardize preclinical assays to measure the potency of such interventions in animal models. Methods Two assay configurations were studied using transgenic Plasmodium berghei expressing Plasmodium falciparum full-length circumsporozoite protein. The assays measured 1) reduction in parasite infection of the liver (liver burden) following an intravenous (i.v) administration of sporozoites and 2) protection from parasitaemia following mosquito bite challenge. Two human CSP mAbs, AB311 and AB317, were compared for their ability to inhibit infection. Multiple independent experiments were conducted to define assay variability and resultant impact on the ability to discriminate differences in mAb functional activity. Results Overall, the assays produced highly consistent results in that all individual experiments showed greater functional activity for AB317 compared to AB311 as calculated by the dose required for 50% inhibition (ID50) as well as the serum concentration required for 50% inhibition (IC50). The data were then used to model experimental designs with adequate statistical power to rigorously screen, compare, and rank order novel anti-CSP mAbs. Conclusion The results indicate that in vivo assays described here can provide reliable information for comparing the functional activity of mAbs. The results also provide guidance regarding selection of the appropriate experimental design, dose selection, and group sizes.

scholarly journals High-Throughput Screening by Mass Spectrometry: Comparison with the Scintillation Proximity Assay with a Focused-File Screen of AKT1/PKBα

2007 ◽  
Vol 12 (4) ◽  
pp. 473-480 ◽  
Author(s):  
Andrea K. Quercia ◽  
William A. Lamarr ◽  
Jayhyuk Myung ◽  
Can C. Özbal ◽  
James A. Landro ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
High Throughput Screening ◽  
Kinase Inhibitors ◽  
Label Free ◽  
False Negatives ◽  
Scintillation Proximity Assay ◽  
Therapy Target ◽  
Hit Identification ◽  
Selection Of

Mass spectrometry is an emerging format for label-free high-throughput screening. The main limitation of mass spectrometry is throughput, due to the requirement to purify samples prior to ionization. Here the authors compare an automated high-throughput mass spectrometry (HTMS) system (RapidFire™) with the scintillation proximity assay (SPA). The cancer therapy target AKT1/PKBα was screened against a focused library of kinase inhibitors and IC50 values determined for all compounds that exhibit > 50% inhibition. A selection of additional compounds that exhibited ≤ 50% inhibition in the primary screen was chosen as controls to confirm inactives. The selection of compounds is expected to identify common actives, common inactives, false positives, and false negatives. Agreement is found between HTMS and SPA in terms of primary hit identification and hit confirmation. ( Journal of Biomolecular Screening 2007:473-480)

High-throughput screening and selection of yeast cell lines expressing monoclonal antibodies

2010 ◽  
Vol 37 (9) ◽  
pp. 961-971 ◽  
Author(s):  
Gavin C. Barnard ◽  
Angela R. Kull ◽  
Nathan S. Sharkey ◽  
Seemab S. Shaikh ◽  
Alissa M. Rittenhour ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Yeast Cell ◽  
Cell Lines ◽  
High Throughput ◽  
High Throughput Screening ◽  
Selection Of

scholarly journals Characterization of two in vivo challenge models to measure functional activity of monoclonal antibodies to Plasmodium falciparum circumsporozoite protein

2020 ◽  
Author(s):  
Rama Raghunandan ◽  
Bryan T Mayer ◽  
Yevel Flores-Garcia ◽  
Monica W Gerber ◽  
Raphael Gottardo ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Functional Activity ◽  
Statistical Power ◽  
Rank Order ◽  
Circumsporozoite Protein ◽  
Dose Selection ◽  
New Strategies ◽  
Selection Of

Abstract Background. New strategies are needed to reduce the incidence of malaria, and promising approaches include the development of vaccines and monoclonal antibodies (mAbs) that target the circumsporozoite protein (CSP). To select the best candidates and speed development, it is essential to standardize preclinical assays to measure the potency of such interventions in animal models. Method. Two assay configurations were studied using transgenic P. berghei expressing P. falciparum full-length circumsporozoite protein. The assays measured 1) reduction in parasite infection of the liver (liver burden) following an intravenous (i.v) administration of sporozoites and 2) protection from parasitemia following mosquito bite challenge. Two human CSP mAbs, AB311 and AB317, were compared for their ability to inhibit infection. Multiple independent experiments were conducted to define assay variability and resultant impact on the ability to discriminate differences in mAb functional activity.Results. Overall, the assays produced highly consistent results in that all individual experiments showed greater functional activity for AB317 compared to AB311 as calculated by the dose required for 50 % inhibition (ID50) as well as the serum concentration required for 50% inhibition (IC50). The data were then used to model experimental designs with adequate statistical power to rigorously screen, compare, and rank order novel anti-CSP mAbs. Conclusion. Our results indicate that in vivo assays described here can provide reliable information for comparing the functional activity of mAbs. Our results also provide guidance regarding selection of the appropriate experimental design, dose selection, and group sizes.

scholarly journals Accelerated Formulation Development of Monoclonal Antibodies (mAbs) and mAb-Based Modalities

2015 ◽  
Vol 20 (4) ◽  
pp. 468-483 ◽  
Author(s):  
Vladimir I. Razinkov ◽  
Michael J. Treuheit ◽  
Gerald W. Becker
Keyword(s):  
Monoclonal Antibodies ◽  
High Throughput ◽  
Drug Product ◽  
Storage Conditions ◽  
Formulation Development ◽  
Long Term Storage ◽  
Therapeutic Monoclonal Antibodies ◽  
Term Storage ◽  
Selection Of

More therapeutic monoclonal antibodies and antibody-based modalities are in development today than ever before, and a faster and more accurate drug discovery process will ensure that the number of candidates coming to the biopharmaceutical pipeline will increase in the future. The process of drug product development and, specifically, formulation development is a critical bottleneck on the way from candidate selection to fully commercialized medicines. This article reviews the latest advances in methods of formulation screening, which allow not only the high-throughput selection of the most suitable formulation but also the prediction of stability properties under manufacturing and long-term storage conditions. We describe how the combination of automation technologies and high-throughput assays creates the opportunity to streamline the formulation development process starting from early preformulation screening through to commercial formulation development. The application of quality by design (QbD) concepts and modern statistical tools are also shown here to be very effective in accelerated formulation development of both typical antibodies and complex modalities derived from them.

scholarly journals In vitro and in vivo inhibition of malaria parasite infection by monoclonal antibodies against Plasmodium falciparum circumsporozoite protein (CSP)

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Merricka C. Livingstone ◽  
Alexis A. Bitzer ◽  
Alish Giri ◽  
Kun Luo ◽  
Rajeshwer S. Sankhala ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Monoclonal Antibodies ◽  
Disease Burden ◽  
Vaccine Candidate ◽  
Specific Binding ◽  
Circumsporozoite Protein ◽  
Target Epitope ◽  
Selection Of

AbstractPlasmodium falciparum malaria contributes to a significant global disease burden. Circumsporozoite protein (CSP), the most abundant sporozoite stage antigen, is a prime vaccine candidate. Inhibitory monoclonal antibodies (mAbs) against CSP map to either a short junctional sequence or the central (NPNA)n repeat region. We compared in vitro and in vivo activities of six CSP-specific mAbs derived from human recipients of a recombinant CSP vaccine RTS,S/AS01 (mAbs 317 and 311); an irradiated whole sporozoite vaccine PfSPZ (mAbs CIS43 and MGG4); or individuals exposed to malaria (mAbs 580 and 663). RTS,S mAb 317 that specifically binds the (NPNA)n epitope, had the highest affinity and it elicited the best sterile protection in mice. The most potent inhibitor of sporozoite invasion in vitro was mAb CIS43 which shows dual-specific binding to the junctional sequence and (NPNA)n. In vivo mouse protection was associated with the mAb reactivity to the NANPx6 peptide, the in vitro inhibition of sporozoite invasion activity, and kinetic parameters measured using intact mAbs or their Fab fragments. Buried surface area between mAb and its target epitope was also associated with in vivo protection. Association and disconnects between in vitro and in vivo readouts has important implications for the design and down-selection of the next generation of CSP based interventions.

scholarly journals Development of a Robust High-Throughput Screening Platform for Inhibitors of the Striatal-Enriched Tyrosine Phosphatase (STEP)

2021 ◽  
Vol 22 (9) ◽  
pp. 4417
Author(s):  
Lester J Lambert ◽  
Stefan Grotegut ◽  
Maria Celeridad ◽  
Palak Gosalia ◽  
Laurent JS De Backer ◽  
...  
Keyword(s):  
Drug Discovery ◽  
High Throughput ◽  
Tyrosine Kinases ◽  
High Throughput Screening ◽  
Kinase Inhibitors ◽  
Tyrosine Phosphatase ◽  
Protein Tyrosine Phosphatases ◽  
Synaptic Function ◽  
Label Free ◽  
Protein Tyrosine

Many human diseases are the result of abnormal expression or activation of protein tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs). Not surprisingly, more than 30 tyrosine kinase inhibitors (TKIs) are currently in clinical use and provide unique treatment options for many patients. PTPs on the other hand have long been regarded as “undruggable” and only recently have gained increased attention in drug discovery. Striatal-enriched tyrosine phosphatase (STEP) is a neuron-specific PTP that is overactive in Alzheimer’s disease (AD) and other neurodegenerative and neuropsychiatric disorders, including Parkinson’s disease, schizophrenia, and fragile X syndrome. An emergent model suggests that the increase in STEP activity interferes with synaptic function and contributes to the characteristic cognitive and behavioral deficits present in these diseases. Prior efforts to generate STEP inhibitors with properties that warrant clinical development have largely failed. To identify novel STEP inhibitor scaffolds, we developed a biophysical, label-free high-throughput screening (HTS) platform based on the protein thermal shift (PTS) technology. In contrast to conventional HTS using STEP enzymatic assays, we found the PTS platform highly robust and capable of identifying true hits with confirmed STEP inhibitory activity and selectivity. This new platform promises to greatly advance STEP drug discovery and should be applicable to other PTP targets.

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