The Abrolhos Bank harbors the richest coral reef ecosystem in the South Atlantic Ocean. It exhibits unique geomorphologic structures, is localized in shallow depths, and is divided into two reef regions with an inner arc close to the coast (3–20 m depth) and an outer deeper arc (5–30 m depth). This study aims to describe some bio-optical properties of the Abrolhos Bank waters and to evaluate the performance of the inversion Hyperspectral Optimization Processing Exemplar (HOPE) model, developed to retrieve optical properties in shallow waters, in the region. To this end, measurements at 75 stations during two field campaigns conducted during the 2013 and 2016 wet seasons were analyzed, and the HOPE model was applied to both in situ remote sensing reflectance (Rrs) spectra and PRecursore IperSpettrale della Missione Applicativa (PRISMA) imagery. Significant differences in optical and biological properties were found between the two arcs. The empirical relationships between chlorophyll-a concentration (Chl-a) and absorption coefficient of phytoplankton at 440 nm (aphy(440)) diverged from Bricaud’s models, suggesting differences in phytoplankton diversity and cell size. In both arcs, total non-water absorption coefficient at 440 nm (aT-w(440)) was dominated by colored dissolved organic matter (CDOM) by ∼60%. Absorption coefficient by CDOM (acdom) presented a higher variability within the outer arc, with the lowest contribution from non-algal particles (NAPs), and the spectral slopes of aCDOM resembled those of the inner arc. The spectral slopes of the NAP absorption coefficient suggested a dominance by organic rather than mineral particles that probably originated from biological production. The HOPE model applied to in situ Rrs performed satisfactorily for depth in the Abrolhos Bank waters, although retrievals of aphy(440), CDOM plus NAP (adg(440)) and aT-w(440) were underestimated with a relative bias of −27.9%, −32.1% and −45.8%, respectively. The HOPE model retrievals from the PRISMA image exhibited low aphy(440) values over the whole scene and the highest adg(440) values in the Caravelas river plume. Very shallow depths (≤3 m), bottom substrate reflectance used as input in the HOPE model, model parametrization associated with the water complexity in the study site, and uncertainties associated to Rrs measurements used as input might be responsible for differences found when comparing HOPE retrievals with in situ measurements.