Evidence map›Paper›PMID 34859053›Full record

ArticleFrontiers in molecular biosciences2021

Monolithic Papain-Immobilized Enzyme Reactors for Automated Structural Characterization of Monoclonal Antibodies.

Francesca Rinaldi, Sara Tengattini, Gloria Brusotti, Giuseppe Tripodo, Benjamin Peters, Caterina Temporini, Gabriella Massolini, Enrica Calleri

Open access · goldAbstract read
In one paragraph

Article in Frontiers in molecular biosciences, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
0.6field-weighted citation impact, top 36% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

2 citing papers in PubMed, 8 citations in OpenAlex.

  1. Article
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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

8 authors at 2 institutions in 2 countries.

Francesca RinaldiDepartment of Drug Sciences, University of Pavia, Pavia, Italy.
Sara TengattiniDepartment of Drug Sciences, University of Pavia, Pavia, Italy.
Gloria BrusottiDepartment of Drug Sciences, University of Pavia, Pavia, Italy.
Giuseppe TripodoDepartment of Drug Sciences, University of Pavia, Pavia, Italy.
Benjamin PetersInstrumental Analytics R&D, Merck KGaA, Darmstadt, Germany.
Caterina TemporiniDepartment of Drug Sciences, University of Pavia, Pavia, Italy.
Gabriella MassoliniDepartment of Drug Sciences, University of Pavia, Pavia, Italy.
Enrica CalleriDepartment of Drug Sciences, University of Pavia, Pavia, Italy.
University of Pavia · ITMerck (Germany) · DE

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

The characterization of monoclonal antibodies (mAbs) requires laborious and time-consuming sample preparation steps before the liquid chromatography-mass spectrometry (LC-MS) analysis. Middle-up approaches entailing the use of specific proteases (papain, IdeS, etc.) emerged as practical and informative methods for mAb characterization. This work reports the development of immobilized enzyme reactors (IMERs) based on papain able to support mAb analytical characterization. Two monolithic IMERs were prepared by the covalent immobilization of papain on different supports, both functionalized via epoxy groups: a Chromolith® WP 300 Epoxy silica column from Merck KGaA and a polymerized high internal phase emulsion (polyHIPE) material synthesized by our research group. The two bioreactors were included in an in-flow system and characterized in terms of immobilization yield, kinetics, activity, and stability using Nα-benzoyl-L-arginine ethyl ester (BAEE) as a standard substrate. Moreover, the two bioreactors were tested toward a standard mAb, namely, rituximab (RTX). An on-line platform for mAb sample preparation and analysis with minimal operator manipulation was developed with both IMERs, allowing to reduce enzyme consumption and to improve repeatability compared to in-batch reactions. The site-specificity of papain was maintained after its immobilization on silica and polyHIPE monolithic supports, and the two IMERs were successfully applied to RTX digestion for its structural characterization by LC-MS. The main pros and cons of the two supports for the present application were described.

Indexed as

analytical characterizationimmobilized enzyme reactor (IMER)LC-MSmiddle-up analysismonoclonal antibodiesmonolithic supportspapainpolymerized high internal phase emulsions (polyHIPEs)

Identifiers

PMID34859053
PMCPMC8630785
OpenAlexW3212163085

What OpenQuestion holds

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LicenceCC BY
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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.