Due to the advantages of microfluidic devices in the design of diagnostic methods, the integration of microfluidic-based devices with iLAMP increases its applicability in the clinical area. The scheme of microfluidic chip-based iLAMP platform is depicted in Figure 4. Figure 4 Integration of microfluidics

with iLAMP (microfluidics-iLAMP platform). Integration with aptamer (aptamer-iLAMP) One of the challenges of current nucleic acid-based detection of proteins is the availability of antibody-signal DNA conjugates. In practice, the preparation of such conjugates is challenging, GDC-0449 in vivo and sometimes the produced conjugates do not have the required specificity and produce background noise [20]. Due to the application of such conjugates in iLAMP, this problem also remains in iLAMP method. However, this

problem can be solved by application of aptamers, the nucleic acids with the ability of specific recognition of their target molecules in the folded conformation instead of antibodies in iLAMP. Beside this advantage, aptamers selleck have many benefits over antibodies. These advantages include low cost, ease of preparation, high stability, the possibility of reversible denaturing, the possibility of on-demand changes of the properties, the possibility of aptamer identification for toxins as well as molecules that do not elicit good immune responses, the minimum batch-to-batch difference in the performance, and the possibility of precise conjugation

with various reporter molecules [60, 61]. Aptamers also have high sensitivity and specificity toward their targets. They have the equilibrium dissociation constants (Kd) of the pico- to micromolar range and can discriminate subtle differences in the structure of their targets, even better than antibodies [62, 63], while some disadvantages can limit the application of antibodies. Laborious production, limited shelf-life, restriction in the production of different types of antibodies due to the limitations in the growth of some hybridomas, Protein kinase N1 batch-to-batch differences in the performance of the same antibody, and inability to modify the kinetic parameters of antibody-target interactions are the main drawbacks of the use of antibodies [63]. Based on the advantages of aptamers over antibodies, aptamer-iLAMP method can be considered as an improved HSP phosphorylation configuration of iLAMP technique. In addition, the aptamers used can serve as both recognition and signal molecule for direct detection of the target without need for antibody-DNA conjugates (Figure 5). Figure 5 Application of aptamer instead of antibody in iLAMP (aptamer-iLAMP platform). Possible limitations of iLAMP and their solutions Like any new detection method, iLAMP may have some potential problems. One problem can be the challenging preparation of antibody-DNA conjugates.