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Peptides not only compose proteins but also are extremely important endogenous compounds playing important roles in living systems.Benefiting from the revolutionary concept of ‘‘lab on a chip,the synthesis and screening of bioactive peptides can be significantly miniaturized,automated and integrated.In our study,a novel integrated continuous-flow microfluidic system was designed and fabricated for solid phase peptide synthesis.The transfer losses,hazardous handling and time-consuming processes in conventional peptide synthesis were avoided by being replaced with the on-chip cleavage treatment.Six peptides against beta-endorphin antibody were obtained simultaneously with high purity within a short time.Furthermore,a novel integrated tetra-layer microfluidic hybrid chip with 36 screening chambers was designed and fabricated for affinity peptide screening with in situ detection.The combination of "reverse" affinity chromatographic assay with the microfluidic technique facilitated multi parameter screening and significantly simplified reagent delivery procedure in a micro-scale.By using six pH gradient generators,the magnetic bead-based screening in parallel chambers was simultaneously carried out under 6 different pH conditions from 5.4 to 8.2.A pentapeptide YGGFL was in situ screened out with the highest affinity towards beta-endorphin antibody at pH 7.1.Our microfluidic based solid phase synthesis and screening systems are automatic,low-cost and recyclable with tremendously practical significance not only for peptide screening but also to other biomedical and bioanalytical affinity researches towards target molecules.