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Case report:
Client
DMC Biotechnologies
Sector
Research
Research Domain
Detection tool development
Target
Engineered DAS+4 tag
Key processes:
- Hybridoma development against peptides
- ELISA screenings to improve coating and presentation
- Antibody production and quality control
Key Numbers
0
High-performing clones developed
0mg
Purified monoclonal antibody produced
Context
Dynamic Metabolic Control (DMC) has developed a metabolic engineering platform enabling the on-demand downregulation of specific pathway enzymes through targeted proteolytic degradation. This is achieved by appending a short synthetic peptide sequence, the DAS+4 tag, directly to selected enzymes, marking them for controlled degradation in response to defined process conditions.
To support the development and optimization of this platform, a detection tool capable of specifically recognizing the DAS+4 tag was required. A key challenge resided in achieving high selectivity against this synthetic sequence while avoiding cross-reactivity with closely related native bacterial sequences. The generation of an anti-DAS+4 antibody with such discriminating specificity would enable the quantification of tagging efficiency and degradation kinetics using standard immunoanalytical techniques, including Western blot and ELISA, thereby providing a reliable analytical framework for strain characterization and bioprocess development.
Challenges
1
High selectivity avoiding cross-reactivity with closely related native bacterial sequences
2
Ensuring functional performance in Western blot and ELISA assays
ProteoGenix Approach
Step I: Peptide design & synthesis
Based on the DAS+4 tag sequence, up to 3 candidate peptides were designed and synthesized, then conjugated to carrier proteins (KLH for immunization, BSA for ELISA screening).
Step II: Mice immunizations
Five mice were immunized using an optimized fast-track protocol with the proprietary TiterFast™ adjuvant. Immune response was monitored by serial bleedings and ELISA titration against the peptide mix.
Step III: Fusion and screenings
Spleen cells from the best-responding mouse were fused with a myeloma cell line. Resulting hybridomas were screened by ELISA to identify clones specifically recognizing the DAS+4 peptides, with up to 20 candidate supernatants shipped for client validation.
Step IV: Hybridoma sub-cloning
The two best parental clones were sub-cloned through iterative rounds of limiting dilution and ELISA screening. Isotyping was performed, and the best monoclonal hybridoma was cryopreserved.
Step V: Antibody production
2 mg of purified monoclonal antibody was produced from the selected clone, purified by Protein A/G affinity chromatography, and quality-controlled by SDS-PAGE and ELISA.
Deliverables:
- Up to 20 best clones (supernatant samples) at polyclonal stage for customer’s selection
- Final monoclones supernatant samples to confirm hybridoma
- 1 hybridoma cell line
- 2 mg of purified antibody
- 2 mg of peptides mix antigen
- Project report
- Ownership and intellectual property of the antibodies
Results
The selected antibody showed strong ELISA reactivity against the targeted peptide (OD 2.95 at 1:2000), while remaining near background on the negative-control peptide (maximum at OD 0.04 across all dilutions). Specific signal was still detectable at 1:128000 dilution. The target signal is about 14.3-fold higher than the control even at the highest dilution tested, supporting high selectivity for the engineered DAS+4 tag.
Key Takeway
ProteoGenix helped DMC Biotechnologies develop a selective antibody against the engineered DAS+4 tag, with strong ELISA signal on the target peptide, minimal reactivity to the similar native-sequence control, and activity maintained over a wide dilution range. Through tailored peptide design, counter-screening, and guided clone selection, the collaboration provided DMC with a practical tool to monitor tag detection and support strain development efforts.
ProteoGenix has been an outstanding partner, with a customer service team that is responsive, knowledgeable, and genuinely invested in our projects.
The delivered antibody consistently shows excellent sensitivity and specificity, delivering robust, reproducible signals with minimal background across both ELISA and Western blot applications.
Combined with very competitive pricing and staying true to timeline obligations, ProteoGenix offers exceptional value, and we highly recommend them to any lab seeking reliable, bespoke, high-quality antibodies.
Collaboration
The success of this collaboration came from a joint effort supported by close communication, scientific expertise, and dedicated management from start to finish. For DMC Biotechnologies, this meant not only access to an IP-free antibody fully owned by the client, but also the reassurance of a fully guaranteed no win, no fee service that helped reduce project risk. With a dedicated PhD expert guiding the project and end-to-end support throughout development and validation, the collaboration provided a strong framework to advance a selective antibody tailored to DMC’s needs.
DMC Biotechnologies
DMC Biotechnologies is a U.S.-based biomanufacturer specializing in precision fermentation and sustainable chemical production. Its proprietary Dynamic Metabolic Control™ platform is built to deliver more standardized, robust, and predictable fermentation processes, with an emphasis on scalable development and production. This work directly supported DMC’s platform by enabling selective detection of the engineered DAS+4 tag used to control enzyme degradation.
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