2022 AMI Online Salon

Sequencing-Linked Immuno-sorbent Assay

Project Details

  • Entrant Name:  Shirley Li
  • Client: Sandra B. Gabelli, Fabian de Kok-Mercado
  • Copyright:  Shirley Li, 2021
  • Medium/software used: Adobe Illustrator, photoshop, and Cinema 4D
  • Final presentation format: half page, journal article
  • Primary Audience: Molecular biologists/immunologists

Project Description

This illustration summarizes key steps of a Sequencing-Linked Immuno-sorbent Assay (SLISY) for generation of antibodies targeting SARS-CoV-2. SLISY is a novel method that uses sequencing to simultaneously assess the entire sequence diversity within a selected phage pool for identification of clones with high specificity. This method had never been visualized in the past. In step 1, a single-chain variable fragment (scFv) phage library is negatively selected against the MERS spike protein, followed by rounds of positive selection against the SARS-CoV-2 spike protein depicted in step 2. In step 3, the complementarity-determining region H3 (CDRH3) region of the scFv is sequenced; a SLISY ratio is determined for each clone, representing the specificity to SARS-CoV-2. The entire scFv region of highly specific clones are sequenced in step 4. The gene block is then cloned into phagemid vectors to express monoclonal scFv, as shown in step 5. Finally, in step 6, monoclonal antibodies are generated for a validation study. In this illustration, arrows were used to help guiding viewers through the flow of the story. Simplified, repetitive elements were used to highlight the core action in each step, which provides an easy-to-follow visualization of this complex method involving multiple scales.