PXD073800-1

PXD073800 is an original dataset announced via ProteomeXchange.

Title	From Sea to Therapy: Development and analytical control of recombinant human CDKL5 production in the marine bacterium Pseudoalteromonas haloplanktis TAC125
Description	Marine bacteria are increasingly explored as alternative microbial platforms for the production of high-value biopharmaceuticals, owing to their distinctive physiological and biochemical features. In this study, we focus on the Antarctic marine bacterium Pseudoalteromonas haloplanktis TAC125 (PhTAC125), an unconventional host that has proven capable of producing soluble and active human Cyclin-Dependent Kinase-Like 5 (hCDKL5). This serine/threonine kinase is essential for neuronal development as its deficiency causes CDKL5 Deficiency Disorder, a severe and currently untreatable neurodevelopmental disease. Recombinant production of hCDKL5 represents a necessary prerequisite for the development of enzyme replacement therapy as a potential therapeutic strategy; however, the current production process remains insufficient for industrial translation. To address this limitation, we developed and implemented dedicated analytical methods to assess both the quantity and functional quality of recombinant hCDKL5. Production yields were evaluated using a customised quantitative sandwich Enzyme-Linked Immunosorbent Assay (ELISA) specifically detecting full-length hCDKL5, while protein functionality was assessed by mass spectrometry-based quantification of autophosphorylation levels, a key indicator of kinase activity. These tools were applied to characterise hCDKL5 expression under different growth conditions. Overall, this work establishes an integrated analytical framework aligned with a Quality by Design approach, providing a robust basis for future process optimisation toward scalable hCDKL5 production.
HostingRepository	PRIDE
AnnounceDate	2026-05-29
AnnouncementXML	Submission_2026-05-29_05:49:43.547.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	ilaria iacobucci
SpeciesList	scientific name: Pseudoalteromonas haloplanktis; NCBI TaxID: NEWT:228;
ModificationList	phosphorylated residue
Instrument	Orbitrap Exploris 240

Revision	Datetime	Status	ChangeLog Entry
0	2026-01-29 14:13:38	ID requested
⏵ 1	2026-05-29 05:49:44	announced

Coletti A, Calvanese M, Cozzolino F, Iacobucci I, Lauro C, Severino A, Monti M, Parrilli E, Tutino ML, TAC125. Mar Drugs, 24(5):(2026) [pubmed]

10.3390/md24050151;

submitter keyword: Antarctic Marine Bacteria

Cell-factory

Difficult to express Proteins

ELISA

hCDKL5

Protein Phosphorylation

Pseudoalteromonas haloplanktis TAC125

Quality by Design.

flora cozzolino
contact affiliation	University of Naples Federico II and CEINGE Biotecnoloige Avanzate "Franco Salvatore"
contact email	flora.cozzolino@unina.it
lab head
ilaria iacobucci
contact affiliation	Department of Chemical Sciences, University of Naples Federico II, Naples 80126, Italy CEINGE Advanced Biotechnologies, University of Naples Federico II, Naples 80145, Italy
contact email	iacobucci@ceinge.unina.it
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD073800

PRIDE project URI

• PRIDE
  1. PXD073800
     1. Label: PRIDE project
     2. Name: From Sea to Therapy: Development and analytical control of recombinant human CDKL5 production in the marine bacterium Pseudoalteromonas haloplanktis TAC125