CamPSF workflow: selections by phage and yeast display

Modular approach taken enabling flexibility for users depending on expertise

Part 1: Pre-selection preparation

a)  Advice on the antigen requirements, its production and optional biotinylation (assistance might also be a possibility), and choosing which libraries will be used – one of ours or other custom libraries supplied by the user (can be non-antibody related scaffolds etc), and planning selections.

 b)  Assistance in writing grant proposal to secure funding to carry out selections  (cost- recovery not for profit model to make the facility self-sustaining with a sliding scale of costs depending on how much work facility carries out

 c)  Users can be actively involved in selections with students/PDRAs trained in techniques and equipment; or can be fully run as a service with user simply providing the target (antigen)

Part 2a: Selections – phage display

CamPSF performs selection of binders using primarily phage display, which allows for a high diversity of sequences (~billions to 10's of billions). Selections target :

• soluble proteins (with additional bound ligands) 
• purified membrane proteins (with additional bound ligands)

• paired cell lines where one displays the surface antigen and one where it is deleted

  In vitro based selections allow precise control of the conditions where binding selections occur, a distinct advantage over animal based immunization approaches

   

Part 2b: Selections – optional yeast display

Enriched library can be transferred to yeast display, which accommodates a reduced diversity (tens of millions of sequences) but can be followed by Magnetic Activated Cell  Sorting(MACS)/Fluorescence Activated Cell Sorting (FACS) and allows a quantitative analysis of affinity as well as multi-factor sorting - currently in development in facility.