In this study, a new method was developed to successfully design sustainable microfibers from wheat gluten proteins using a nonreducing solvent and electrospinning. We explored the morphology by X-ray tomography, scanning electron microscopy (SEM), and confocal laser scanning microscopy (CLSM), protein chemistry and cross-linking by size exclusion-high-performance liquid chromatography (SE-HPLC), and secondary structure by Fourier transform infrared spectroscopy (FT-IR) of fibers containing 15 and 20% of gluten. The impact of heat (130 °C) post-treatment on the polymerization properties of fibers and their absorption performance in different biofluids were also evaluated. The fibers with 20% gluten showed a uniform architecture supported by a relatively stronger fibrous network as compared to irregular and brittle fibers from 15% gluten. Heat treatment of fibers increased the protein cross-linking in all electrospun fibers as compared to the non-heat-treated fibers, as evidenced by SE-HPLC. An increase in the amount of α-helices and random coils was observed in the proteins of all of the heat-treated fibers compared to the nontreated fibers by FT-IR. This suggested that the heat treatment contributed positively to the gluten protein's chemical rearrangements, e.g., aggregation, new hydrogen and isopeptide bonding, and conversion of some of the sulfhydryl groups into disulfide cross-links, contributing positively to the functional performance. The heat-treated electrospun fibers with 20% gluten showed a very attractive blood absorption capacity (323%) and reasonable stability in phosphate-buffered saline (PBS) buffer compared to 15% gluten fibers and non-heat-treated fibers. Cotton-like fiber architecture, high blood absorption capacity, and reasonable stability in PBS buffer are properties desired for absorbents of biofluids and should be further explored in healthcare and medical applications.