Site Specific Fluorescent Labeling via SpyTag SpyCatcher for Rapid Hybridoma Screening in Semi-Solid Medium
Guo, A.; Wei, M.; Wu, J.; Li, X.; Jiang, B.
Show abstract
Hybridoma screening in semi-solid medium typically employs antigens labeled with visible fluorophores (e.g., FITC, AF488) to enable single-step identification of antibody-secreting clones. However, conventional chemical conjugation via NHS-esters or isothiocyanate groups frequently modifies lysine residues located within epitopes, potentially abrogating antibody recognition of these critical regions. Here, we describe a SpyTag SpyCatcher-based site-specific labeling strategy that circumvents epitope damage during semi-solid medium screening. A 16-amino-acid SpyTag was genetically fused to the C-terminus of the target antigen, enabling covalent conjugation to an sfGFP SpyCatcher fluorescent probe. In semi-solid medium supplemented with SpyTag-antigen and sfGFPSpyCatcher, positive hybridoma clones were readily identified by distinct fluorescent halos, whereas negative clones showed no detectable signal. Notably, the site-specific method yielded a significantly higher frequency of fluorescence-positive clones compared to the conventional AF488-labeled antigen method, suggesting that epitope preservation enhances screening recovery. Furthermore, this approach did not impair hybridoma growth or final clone positivity, offering a simple, rapid, and epitope-compatible method for monoclonal antibody screening.
Matching journals
The top 8 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- Enhancement of antibody thermostability and affinity by computational design in the absence of antigen 93%
- A high-throughput platform for biophysical antibody developability assessment to enable AI/ML model training 91%
- A novel biparatopic antibody-ACE2 fusion that blocks SARS-CoV-2 infection: implications for therapy 90%
Similar papers in this journal
- Accelerated Antibody Discovery Targeting the SARS-CoV-2 Spike Protein for COVID-19 Therapeutic Potential 92%
- Broadly Reactive Anti-VHH Antibodies for Characterizing, Blocking, or Activating Nanobody-Based CAR-T Cells 91%
- A Rapid Bead-Based Assay For Screening Of SARS-CoV-2 Neutralising Antibodies 90%
Similar papers in this journal
- Shrimp Hemocyanin elicits a potent humoral response in mammals and is favorable to hapten conjugation 91%
- Rapid functionalisation and detection of viruses via a novel Ca2+-mediated virus-DNA interaction 91%
- Giant magnetoresistive biosensors for real-time quantitative detection of protease activity 91%
Similar papers in this journal
- Engineered Reversible Inhibition of SpyCatcher Reactivity Enables Rapid Generation of Bispecific Antibodies 92%
- Multiplex secretome engineering enhances recombinant protein production and purity 91%
- SARS-CoV-2 neutralizing human recombinant antibodies selected from pre-pandemic healthy donors binding at RBD-ACE2 interface 91%
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.