Evidence map›Paper›PMID 40259001›Full record

ReviewNature protocols2025

Direct analysis of biotransformations with mass spectrometry-DiBT-MS.

Ruth Knox, Rachel Smith, Emily E Kempa, Reynard Spiess, Christian Schnepel, Nicholas J Turner, Sabine L Flitsch, Perdita E Barran

Abstract readReview
PubMed Publisher
In one paragraph

Review in Nature protocols, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.

0numbers the graph read from it
0cells of the map it votes in
3citing papers in PubMed
–field-weighted citation impact
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

3 citing papers in PubMed.

  1. Next-Generation Manufacturing: The Evolving Role of Biocatalysis in AstraZeneca.Chembiochem : a European journal of chemical biology · 2026
    Review
  2. Article
  3. Article
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.

Ruth Knox *Michael Barber Centre for Collaborative Mass Spectrometry, Department of Chemistry, Manchester Institute of Biotechnology, University of Manchester, Manchester, UK.
Rachel Smith *Michael Barber Centre for Collaborative Mass Spectrometry, Department of Chemistry, Manchester Institute of Biotechnology, University of Manchester, Manchester, UK.ORCID 0000-0002-4665-7003
Emily E KempaMichael Barber Centre for Collaborative Mass Spectrometry, Department of Chemistry, Manchester Institute of Biotechnology, University of Manchester, Manchester, UK.ORCID 0000-0001-6481-7974
Reynard SpiessMichael Barber Centre for Collaborative Mass Spectrometry, Department of Chemistry, Manchester Institute of Biotechnology, University of Manchester, Manchester, UK.ORCID 0009-0004-4692-9787
Christian SchnepelMichael Barber Centre for Collaborative Mass Spectrometry, Department of Chemistry, Manchester Institute of Biotechnology, University of Manchester, Manchester, UK.ORCID 0000-0002-4331-2675
Nicholas J TurnerMichael Barber Centre for Collaborative Mass Spectrometry, Department of Chemistry, Manchester Institute of Biotechnology, University of Manchester, Manchester, UK.ORCID 0000-0002-8708-0781
Sabine L FlitschMichael Barber Centre for Collaborative Mass Spectrometry, Department of Chemistry, Manchester Institute of Biotechnology, University of Manchester, Manchester, UK.ORCID 0000-0003-3974-646X
Perdita E BarranMichael Barber Centre for Collaborative Mass Spectrometry, Department of Chemistry, Manchester Institute of Biotechnology, University of Manchester, Manchester, UK. perdita.barran@manchester.ac.uk.ORCID 0000-0002-7720-586X

Funding

RCUK | Biotechnology and Biological Sciences Research Council (BBSRC) BB/M017702/1RCUK | Engineering and Physical Sciences Research Council (EPSRC) EP/S005226/1RCUK | Engineering and Physical Sciences Research Council (EPSRC) EP/T019328/1
6 · The paper itself

Abstract

The development and analysis of engineered enzymes is greatly assisted by the use of high-throughput screening to quickly determine the efficacy of biotransformations under various conditions. Ambient ionization, particularly desorption electrospray ionization (DESI), coupled to high-resolution mass spectrometry has the advantages of minimal requirements for sample preparation before analysis, which renders it suitable for high-throughput screening, in which the accurate mass and potentially the tandem mass spectrometry (MS) fingerprint for any given product can be used for identification. We present a protocol that permits the application of this method in routine biotechnology and chemical biology laboratories that are using engineered enzymes (such as imine reductases and carboxylic acid reductases, mentioned herein) to produce target compounds from substrates (quinoline moieties and phenyl(piperazinyl) moieties, respectively). Through the use of DESI's MS imaging capabilities, reaction monitoring can be easily visualized via imaging of selected substrate or product ions in a convenient, user-friendly workflow. We describe here how DESI-MS can be used to directly analyze the activity of biotransformations from crude cell lysate, which we term 'DiBT-MS'. The DiBT-MS method presented here is 10-1,000 times as fast as liquid chromatography-MS, with the full procedure for 96 samples taking ~2 h and consuming far less solvent and sample. Also demonstrated in this protocol is the impact of solvent spray composition on ionization efficiency of the target analyte, the benefits of a nylon membrane slide and the reusability of sample slides in multiple experiments.

Indexed as

BiotransformationSpectrometry, Mass, Electrospray IonizationTandem Mass Spectrometry

Identifiers

What OpenQuestion holds

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Read underepoch 390

Registered trials

None linked

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.