06/03/2022

Challenges in the Development of a Method for the Detection of Anti-PEGylated-aptamer Antibodies

Immunogenicity assays capable of properly determining the presence of anti-drug antibodies (ADA) and anti-PEG antibodies have been recognized of great importance due to the fact that pre-existing anti-PEG antibodies could have an impact on the efficacy of a drug coupled to polyethylene glycol (PEG).

AON-D21 is a clinical stage PEGylated L-configured oligonucleotide aptamer targeting complement factor C5a. In this study, the sponsor required a method to be developed for the determination of anti-AON-D21 and anti-PEG antibodies in human serum.

In the work we present to the 8th EBF Young Scientist Symposium, a number of challenges were faced during the development of the assay mainly around robust detection of positive control antibodies by using biotin and sulfo-tag labelled AON-D21 as capture and detection reagents. Several methods were evaluated with the most optimal assay design being the Affinity Capture Elution- AGL (ACE-AGL) format, which captures the ADA binding to immobilized biotin-AON-D21, followed by an acid elution step of ADA and finally the re-capture of these on a MSD plate coated with protein AGL. The ADA are then detected using sulfo-tag labelled AON-D21. In confirmatory assays, this is competed by unlabelled AON-D21 or PEG alone to identify the ADA binding site.

Special emphasis was put in identifying the best positive control by assessing different monoclonal anti-PEG antibodies in order to optimise assay sensitivity and reproducibility.

Finally, a suitable method capable of detecting both anti-PEG and anti-AON-D21 antibodies simultaneously was developed. The method has been validated and is being used to support a Phase I clinical trial in humans

You may find below the poster for your ready reference:

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26/05/2022 11:39

Challenges in the determination of two analytes with different physicochemical properties, Tapinarof and its main metabolite, Tapinarof Sulfate, at the low picogram level

The challenge of developing a new bioanalytical method considerably increases when two or more metabolites need to be simultaneously analysed. Their extraction and chromatography optimization can be further complicated if their physicochemical properties vary significantly. Method development may be even more difficult when low limits of quantitation are required, for instance, when evaluating the systemic adsorption of topical drugs.

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07/04/2022 10:15

Anapharm Bioanalytics Brazilian ANVISA Certificate Renewal

Anapharm Bioanalytics is proud to announce that it has recently renewed its ANVISA (Brazil) certification which is now valid until February 26th, 2024.

READ MORE
23/03/2022 15:12

Anapharm Bioanalytics acquires a new high sensitivity equipment: SCIEX Triple Quad 6500+ system

Anapharm Bioanalytics’ small molecule division continues to expand its capabilities with the acquisition of a new LC-MS/MS equipment tailored for method development and validation of high sensitivity assays

READ MORE

05/26/2022

Challenges in the determination of two analytes with different physicochemical properties, Tapinarof and its main metabolite, Tapinarof Sulfate, at the low picogram level

The challenge of developing a new bioanalytical method considerably increases when two or more metabolites need to be simultaneously analysed. Their extraction and chromatography optimization can be further complicated if their physicochemical properties vary significantly. Method development may be even more difficult when low limits of quantitation are required, for instance, when evaluating the systemic adsorption of topical drugs.

In the work we present to the 8th EBF Young Scientist Symposium, which was awarded with the "Second Best Poster Award", we show the challenges encountered and the potential solutions of developing a fairly sensitive method, while trying to simultaneously extract and analyse two analytes, Tapinarof and, its main phase-II metabolite, Tapinarof Sulfate, whose physicochemical properties are quite different.

You may find below the poster for your ready reference:

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03/06/2022 12:39

Challenges in the Development of a Method for the Detection of Anti-PEGylated-aptamer Antibodies

Immunogenicity assays capable of properly determining the presence of anti-drug antibodies (ADA) and anti-PEG antibodies have been recognized of great importance due to the fact that pre-existing anti-PEG antibodies could have an impact on the efficacy of a drug coupled to polyethylene glycol (PEG).

READ MORE
07/04/2022 10:15

Anapharm Bioanalytics Brazilian ANVISA Certificate Renewal

Anapharm Bioanalytics is proud to announce that it has recently renewed its ANVISA (Brazil) certification which is now valid until February 26th, 2024.

READ MORE
23/03/2022 15:12

Anapharm Bioanalytics acquires a new high sensitivity equipment: SCIEX Triple Quad 6500+ system

Anapharm Bioanalytics’ small molecule division continues to expand its capabilities with the acquisition of a new LC-MS/MS equipment tailored for method development and validation of high sensitivity assays

READ MORE

04/07/2022

Anapharm Bioanalytics Brazilian ANVISA Certificate Renewal

Anapharm Bioanalytics is proud to announce that it has recently renewed its ANVISA (Brazil) certification which is now valid until February 26th, 2024.

Anapharm has been certified by Brazilian regulatory authority since the first inspection took place in 2011. We have since then been supporting our European and Latin American customers achieving their goals in Brazilian market and will gladly continue our mission of providing high-quality bioanalytical services in the coming years.
Should you have any project that might require our bioanalytical support, please do not hesitate to contact our Business Development team, at contact@anapharmbioanalytics.com.

About Anapharm Bioanalytics

30+ years’ experience in LC-MS/MS Bioanalysis and 15+ years’ experience in LBA assays for biologics, acknowledges Anapharm Bioanalytics as a world class provider of bioanalytical services to international Sponsors with its strategically located, GLP-certified and GCP-compliant laboratory in Barcelona.

With a successful regulatory history, having undergone 22 successful inspections from health agencies, including FDA (14 studies), WHO GCP/GLP compliance, GCP inspections from Austria, Italy and Spain (12 studies), as well as GLP certifications by EU authorities and ANVISA (Brazil), Anapharm has become a reliable analytical partner to support its clients worldwide at any stage of drug development from preclinical throughout all clinical stages Phase I-IV.

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03/06/2022 12:39

Challenges in the Development of a Method for the Detection of Anti-PEGylated-aptamer Antibodies

Immunogenicity assays capable of properly determining the presence of anti-drug antibodies (ADA) and anti-PEG antibodies have been recognized of great importance due to the fact that pre-existing anti-PEG antibodies could have an impact on the efficacy of a drug coupled to polyethylene glycol (PEG).

READ MORE
26/05/2022 11:39

Challenges in the determination of two analytes with different physicochemical properties, Tapinarof and its main metabolite, Tapinarof Sulfate, at the low picogram level

The challenge of developing a new bioanalytical method considerably increases when two or more metabolites need to be simultaneously analysed. Their extraction and chromatography optimization can be further complicated if their physicochemical properties vary significantly. Method development may be even more difficult when low limits of quantitation are required, for instance, when evaluating the systemic adsorption of topical drugs.

READ MORE
23/03/2022 15:12

Anapharm Bioanalytics acquires a new high sensitivity equipment: SCIEX Triple Quad 6500+ system

Anapharm Bioanalytics’ small molecule division continues to expand its capabilities with the acquisition of a new LC-MS/MS equipment tailored for method development and validation of high sensitivity assays

READ MORE

03/23/2022

Anapharm Bioanalytics acquires a new high sensitivity equipment: SCIEX Triple Quad 6500+ system

Anapharm Bioanalytics’ small molecule division continues to expand its capabilities with the acquisition of a new LC-MS/MS equipment tailored for method development and validation of high sensitivity assays. The new instrument increases our capabilities for peptide chemistry as well as for orally inhaled drugs such as for example Glycopyrronium; Indacaterol, Fluticasone, Salmeterol, Budesonide, Tiotropium, etc. in biological fluids.
The Triple Quad 6500+ system features multi-component IonDrive Technology including the IonDrive High Energy Detector+ that delivers revolutionary sensitivity, speed, and performance for the most challenging methods.

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03/06/2022 12:39

Challenges in the Development of a Method for the Detection of Anti-PEGylated-aptamer Antibodies

Immunogenicity assays capable of properly determining the presence of anti-drug antibodies (ADA) and anti-PEG antibodies have been recognized of great importance due to the fact that pre-existing anti-PEG antibodies could have an impact on the efficacy of a drug coupled to polyethylene glycol (PEG).

READ MORE
26/05/2022 11:39

Challenges in the determination of two analytes with different physicochemical properties, Tapinarof and its main metabolite, Tapinarof Sulfate, at the low picogram level

The challenge of developing a new bioanalytical method considerably increases when two or more metabolites need to be simultaneously analysed. Their extraction and chromatography optimization can be further complicated if their physicochemical properties vary significantly. Method development may be even more difficult when low limits of quantitation are required, for instance, when evaluating the systemic adsorption of topical drugs.

READ MORE
07/04/2022 10:15

Anapharm Bioanalytics Brazilian ANVISA Certificate Renewal

Anapharm Bioanalytics is proud to announce that it has recently renewed its ANVISA (Brazil) certification which is now valid until February 26th, 2024.

READ MORE

02/28/2022

New UPLC-MS/MS bioanalytical method for quantification of Glycopyrronium and Indacaterol

We are glad to announce that a new method has been successfully developed and validated for determination of Glycopyrronium and Indacaterol in human plasma by LC/MS/MS with a calibration range of 1-200 pg/mL and 2-250 pg/mL, respectively.

Since both analytes can be quantified in a single run, it allows for reduced volumes of blood required, a faster turnaround time for sample analysis and significant cost advantages.
The method involves a solid-phase extraction procedure. Glycopyrronium, Indacaterol and internal standards are measured by reversed phase ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS).

The development of this assay adds to the long experience of Anapharm with high sensitivity methods for inhaled drugs including Tiotropium, Fluticasone, Salmeterol and Formoterol.

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03/06/2022 12:39

Challenges in the Development of a Method for the Detection of Anti-PEGylated-aptamer Antibodies

Immunogenicity assays capable of properly determining the presence of anti-drug antibodies (ADA) and anti-PEG antibodies have been recognized of great importance due to the fact that pre-existing anti-PEG antibodies could have an impact on the efficacy of a drug coupled to polyethylene glycol (PEG).

READ MORE
26/05/2022 11:39

Challenges in the determination of two analytes with different physicochemical properties, Tapinarof and its main metabolite, Tapinarof Sulfate, at the low picogram level

The challenge of developing a new bioanalytical method considerably increases when two or more metabolites need to be simultaneously analysed. Their extraction and chromatography optimization can be further complicated if their physicochemical properties vary significantly. Method development may be even more difficult when low limits of quantitation are required, for instance, when evaluating the systemic adsorption of topical drugs.

READ MORE
07/04/2022 10:15

Anapharm Bioanalytics Brazilian ANVISA Certificate Renewal

Anapharm Bioanalytics is proud to announce that it has recently renewed its ANVISA (Brazil) certification which is now valid until February 26th, 2024.

READ MORE

12/02/2021

Challenges in the Detection of Metabolic Biomarkers Using a Multi Plex Assay

In the work we present to the 14th EBF Open Symposium, we show a 5 Plex method for the detection of metabolic biomarkers in human plasma was developed using the U-PLEX technology on the MSD platform.

The complexities surrounding this method were the establishment of endogenous QCs for each chosen biomarker and evaluating the potential use and need for buffer QCs vs matrix QCs where applicable. Additional challenges were faced while trying to ascertain a single minimum required dilution to enable accurate quantification of all biomarkers simultaneously, encompassing the different detection ranges and limits for each biomarker.

The method development focused on the challenges of balancing the detection of the 5 biomarkers; ensuring that each could be quantified within its own required range whilst maintaining a method that was fit for purpose for all 5 biomarkers. To enable this balance, the context of use for each biomarker was evaluated and the method adapted as required.

A fit for purpose method was established that met the requirements for the need and context of use. The method was used successfully in the quantification of human samples from a clinical study.

You may find below the poster for your ready reference:

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03/06/2022 12:39

Challenges in the Development of a Method for the Detection of Anti-PEGylated-aptamer Antibodies

Immunogenicity assays capable of properly determining the presence of anti-drug antibodies (ADA) and anti-PEG antibodies have been recognized of great importance due to the fact that pre-existing anti-PEG antibodies could have an impact on the efficacy of a drug coupled to polyethylene glycol (PEG).

READ MORE
26/05/2022 11:39

Challenges in the determination of two analytes with different physicochemical properties, Tapinarof and its main metabolite, Tapinarof Sulfate, at the low picogram level

The challenge of developing a new bioanalytical method considerably increases when two or more metabolites need to be simultaneously analysed. Their extraction and chromatography optimization can be further complicated if their physicochemical properties vary significantly. Method development may be even more difficult when low limits of quantitation are required, for instance, when evaluating the systemic adsorption of topical drugs.

READ MORE
07/04/2022 10:15

Anapharm Bioanalytics Brazilian ANVISA Certificate Renewal

Anapharm Bioanalytics is proud to announce that it has recently renewed its ANVISA (Brazil) certification which is now valid until February 26th, 2024.

READ MORE

Considerations to properly assess drug stability within biological samples

Assessing drug stability during method development and validation is of paramount importance. The concentration of the target analyte must remain unaffected throughout the lifecycle of the samples to ensure the reliability of the assay data. However, a decrease in analyte concentration in a biological fluid is not always due to a lack of stability.

In the work we present to the 14th EBF Open Symposium, we show several factors that can affect analyte concentration and that might lead to erroneous conclusions regarding its stability.

These misleading phenomena are described in the poster and focus on three main points:
• Non-specific adsorption.
• Equilibrium time between erythrocytes and plasma.
• Plasma age.

You may find below the poster for your ready reference:

MORE NEWS

03/06/2022 12:39

Challenges in the Development of a Method for the Detection of Anti-PEGylated-aptamer Antibodies

Immunogenicity assays capable of properly determining the presence of anti-drug antibodies (ADA) and anti-PEG antibodies have been recognized of great importance due to the fact that pre-existing anti-PEG antibodies could have an impact on the efficacy of a drug coupled to polyethylene glycol (PEG).

READ MORE
26/05/2022 11:39

Challenges in the determination of two analytes with different physicochemical properties, Tapinarof and its main metabolite, Tapinarof Sulfate, at the low picogram level

The challenge of developing a new bioanalytical method considerably increases when two or more metabolites need to be simultaneously analysed. Their extraction and chromatography optimization can be further complicated if their physicochemical properties vary significantly. Method development may be even more difficult when low limits of quantitation are required, for instance, when evaluating the systemic adsorption of topical drugs.

READ MORE
07/04/2022 10:15

Anapharm Bioanalytics Brazilian ANVISA Certificate Renewal

Anapharm Bioanalytics is proud to announce that it has recently renewed its ANVISA (Brazil) certification which is now valid until February 26th, 2024.

READ MORE

11/04/2021

Cholecalciferol (Vitamin D3)

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03/06/2022 12:39

Challenges in the Development of a Method for the Detection of Anti-PEGylated-aptamer Antibodies

Immunogenicity assays capable of properly determining the presence of anti-drug antibodies (ADA) and anti-PEG antibodies have been recognized of great importance due to the fact that pre-existing anti-PEG antibodies could have an impact on the efficacy of a drug coupled to polyethylene glycol (PEG).

READ MORE
26/05/2022 11:39

Challenges in the determination of two analytes with different physicochemical properties, Tapinarof and its main metabolite, Tapinarof Sulfate, at the low picogram level

The challenge of developing a new bioanalytical method considerably increases when two or more metabolites need to be simultaneously analysed. Their extraction and chromatography optimization can be further complicated if their physicochemical properties vary significantly. Method development may be even more difficult when low limits of quantitation are required, for instance, when evaluating the systemic adsorption of topical drugs.

READ MORE
07/04/2022 10:15

Anapharm Bioanalytics Brazilian ANVISA Certificate Renewal

Anapharm Bioanalytics is proud to announce that it has recently renewed its ANVISA (Brazil) certification which is now valid until February 26th, 2024.

READ MORE

Mesalazine

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03/06/2022 12:39

Challenges in the Development of a Method for the Detection of Anti-PEGylated-aptamer Antibodies

Immunogenicity assays capable of properly determining the presence of anti-drug antibodies (ADA) and anti-PEG antibodies have been recognized of great importance due to the fact that pre-existing anti-PEG antibodies could have an impact on the efficacy of a drug coupled to polyethylene glycol (PEG).

READ MORE
26/05/2022 11:39

Challenges in the determination of two analytes with different physicochemical properties, Tapinarof and its main metabolite, Tapinarof Sulfate, at the low picogram level

The challenge of developing a new bioanalytical method considerably increases when two or more metabolites need to be simultaneously analysed. Their extraction and chromatography optimization can be further complicated if their physicochemical properties vary significantly. Method development may be even more difficult when low limits of quantitation are required, for instance, when evaluating the systemic adsorption of topical drugs.

READ MORE
07/04/2022 10:15

Anapharm Bioanalytics Brazilian ANVISA Certificate Renewal

Anapharm Bioanalytics is proud to announce that it has recently renewed its ANVISA (Brazil) certification which is now valid until February 26th, 2024.

READ MORE

Tamoxifen Isomers E and Z

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03/06/2022 12:39

Challenges in the Development of a Method for the Detection of Anti-PEGylated-aptamer Antibodies

Immunogenicity assays capable of properly determining the presence of anti-drug antibodies (ADA) and anti-PEG antibodies have been recognized of great importance due to the fact that pre-existing anti-PEG antibodies could have an impact on the efficacy of a drug coupled to polyethylene glycol (PEG).

READ MORE
26/05/2022 11:39

Challenges in the determination of two analytes with different physicochemical properties, Tapinarof and its main metabolite, Tapinarof Sulfate, at the low picogram level

The challenge of developing a new bioanalytical method considerably increases when two or more metabolites need to be simultaneously analysed. Their extraction and chromatography optimization can be further complicated if their physicochemical properties vary significantly. Method development may be even more difficult when low limits of quantitation are required, for instance, when evaluating the systemic adsorption of topical drugs.

READ MORE
07/04/2022 10:15

Anapharm Bioanalytics Brazilian ANVISA Certificate Renewal

Anapharm Bioanalytics is proud to announce that it has recently renewed its ANVISA (Brazil) certification which is now valid until February 26th, 2024.

READ MORE