Monitoring methods | Suitable targets | Advantages | Limitations | References |
---|---|---|---|---|
Dot blotting | Protein | Focus on the candidate aptamers binding phase, which shows the real enrichment in SELEX; Relative ease of performance | Sequence labeling required; Not suitable for the target molecules with the same electrostatic charge as NC membrane used | [34] |
qPCR | Protein & Small molecule | Relative ease of performance | Focus on the elution phase, which could be confused by non-specific eluates; Error of nonspecific amplification; | |
EMSA | Protein | Focus on the candidate aptamers binding phase, which shows the real enrichment in SELEX | Sequence labeling required; Labor- and time-consuming; Not suitable for small molecule targets | [39] |
Gel-shifting | Protein | Focus on the candidate aptamers binding phase, which shows the real enrichment in SELEX; Easy for performance | Detection in none-binding conditions, I.E. electrophoretic buffer solution; Not suitable for small molecule targets | |
ELONA | Protein | Focus on the candidate aptamers binding phase, which shows the real enrichment in SELEX; Relative ease of performance | Sequence labeling required; Labor- and time-consuming; Non-specific binding of candidates to the plate, which confuse the enrichment; Not suitable for small molecule targets | [40] |
Agarose gel analysis | Protein & Small molecule | Relative ease of performance | Focus on the elution phase, which could be confused by non-specific eluates; Error of nonspecific amplification | [41] |
HTS | Protein & Small molecule | Focus on the enrichment of candidate aptamers according to the SELEX rounds | Expensive cost required; Focus on the elution phase, which could confuse by none-specific elution; Time-consuming for sample preparation | |
SPR | Protein & Small molecule | Focus on the candidate aptamers binding phase, which shows the real enrichment in SELEX | Expensive sensor chip required; Labor- and time-consuming | |
UV quantification | Small molecule | Ease of performance | Focus on the elution phase, which could confuse by none-specific elution | [44] |
Fluorescence quantification | Small molecule | Relative ease of performance | Sequence labeled with fluorophore required; Focus on the elution phase, which could confuse by none-specific elution | |
Fluorescence binding assay | Small molecule | Focus on the candidate aptamers binding phase, which shows the real enrichment in SELEX; Relative ease of performance | The autofluorescence of target required | |
Gel-elution assay | Small molecule | Focus on the candidate aptamers binding phase, which shows the real enrichment in SELEX | Target-coupled column required; Labor- and time-consuming | [49] |
GBDM | Protein & Small molecule | Focus on the candidate-target binding phase, which shows the real enrichment in SELEX; Easy for performance without expensive equipment; Suitable for monitoring every selection step during SELEX process | Overnight diffusion required | Optimized in This work |