Application of cold atmospheric-pressure plasma (CAP) for wound treatment and decontamination of food or water often includes the presence of proteins. These proteins contain amino acids with functional groups, such as thiol (SH) groups. Plasma-mediated effects in biological systems involve reactive oxygen and nitrogen species. In this regard, redox signaling is often mediated via thiol groups. The aim of this study was to investigate the influence of a protein and an SH group containing amino acids on the antimicrobial properties of plasmatreated saline solution (0.85% NaCl, w/v) and on the direct CAP efficacy on solid wet agar plates. Plasma treatment of saline solution was realized using an ACdriven pin-to-liquid discharge. After plasma treatment for 10 minutes, the amino acids L-cysteine (contains SH group) or L-alanine (no SH group) or bovine serum albumin (BSA; with approximately 6% cysteine content) were added together with the test microorganism Escherichia coli K-12 (DSM 11250/ NCTC 10538) for an exposure time of up to 60 min. The total viable cell count was determined at appropriate time intervals. A concentration-dependent repeal of the antimicrobial efficacy was determined. Therefore, 0.0025% of BSA did not have any influence, whereas 0.25% of BSA w/v, as well as the tested amino acids, did not result in the inactivation of E. coli. The difference between L-alanine and L-cysteine was negligible, suggesting only a minor effect of the presence of SH groups. Dimerization of L-cysteine was shown by LC/MS analyses, whereas no derivatization of L-alanine was detected. To test the influence of proteins in direct plasma treatment on wet surfaces, E. coli was plated together with BSA on soybean-casein digest agar surface. Another setup was based on agar plates, which contained different concentrations of peptone (a mixture of peptides and amino acids). The agar plates were regularly treated by the argon-driven CAP jet kINPen Med (neoplas tools GmbH, Greifswald, Germany). After overnight incubation, inhibition zones were analyzed. The bacterial growth was independent of protein or peptone content for this direct plasma treatment. Summarizing all, the antibacterial effect of the Plasma Process Polym. 2019;16:e1800164 www.plasma-polymers.com