Large amounts of coexisting contamination in complex biofluid samples impede the quantified veracity of biomarkers, which is the key problem for disease confirmation. Herein, amyloid-like transformed bovine serum albumin inlaid with gold nanoparticles was used as a coating (BGC) on a substrate composed of silicon nanowires (SW; BGC-SW) under ambient conditions. After modification with the recognition group, BGC-SW could serve as an outstanding platform for the selective separation and sensitive detection of biomarkers in complicated biosamples. First, the BGC on SW with a large surface area exhibits excellent adhesion resistance. The attached amounts of contaminations in biofluids were decreased by over 78% compared with native bovine serum albumin (BSA) as the blocking agent. This is because the phase-transformed BSA coating provides stronger interactions with the SW than bare BSA, which results in a tighter attachment and more uniform coverage of the BGC. Furthermore, the gold matrix laid inside the antiadhesive coating ensures simple cross-linking with the recognition groups to selectively capture various biomarkers in complex biofluids and create a gentle release method. Circulating tumor cells (CTCs) were chosen as template biomarkers to verify the application of A-BGC-SW (BGC-SW modified with sgc8-aptamer) in various separation processes of untreated biofluids. The results showed that approximately six cells could be captured from a 1 mL fresh blood sample containing only 10 CTCs. The easy fabrication and excellent antiadhesion property endow A-BGC-SW with great potential in the field of biological separation.