Table 2 Comparison of the advantages and disadvantages of membranome target discovery techniques
Protein profiling using LPI™ FlowCells and LC-MS/MS | Hybridoma-based phenotypic antibody screening | Phage display-based phenotypic antibody screening | |||
|---|---|---|---|---|---|
Advantages | Disadvantages | Advantages | Disadvantages | Advantages | Disadvantages |
Direct comparison with normal matched tissue can be performed | Extremely reliant on quality of membrane preparation and extraction of membrane proteins | Screening antibodies can assign a potential mechanism of action | High cell requirement | Reduced cell requirement compared to hybridoma approach | Isolation of relatively low affinity antibodies and poor target identification success rate- requirement for complementary techniques for success of target identification |
Sample fractionation possible and total survey of the membranome | Reliant on database annotations | Isolation of high affinity antibodies | Dominance of single targets and antibodies | Ability to perform initial screen against multiple cell types with relative ease | |
No function or mechanism of action associated with antigens identified | Functional in phenotypic screens | No ability to deselect against abundant antigens or comparator cell types | Screening can assign a potential mechanism of action | ||
High target identification success rate | Ability to avoid dominance – can deselect against abundant antigens and comparator cell types | ||||
Isolation of antibodies that can be used for target validation or as a therapeutic candidates | Isolation of antibodies that can be used for target validation | ||||
Potential to identify target and therapeutic candidate | |||||