▎ 摘　　要

Derivatization of 2D materials for bioapplications is at the forefront of nanomaterials research nowadays. Facile synthesis of the biografted 2D derivatives and insight into the conformation of the conjugated biomolecules are two pillars, promoting advances in the field of biosensing, drug delivery and regeneration techniques. This work is devoted to the synthesis and conjugation of carboxylated reduced graphene oxide (C-xy rGO) by aptamers followed by theoretical analysis of their conformation in the immobilized state. Employing the developed method, the hole-matrixed graphene with up to 11.1 at. % reactive carboxyl groups was synthesized and thoroughly examined via core-level spectroscopy. The mechanism of the performed carboxylation with conversion of graphene oxide into carboxylated graphene is proposed, unveiling commonly disregarded impact of ether-like components to the fingerprints of the carboxyl groups. We show successful covalent immobilization of the AO-01 aptamer against Hepatitis B protein on the synthesized C-xy_rGO and for the first time reveal its conformation both in free and immobilized forms via a combination of density functional tight binding (DFTB) calculations and molecular dynamic (MD) modeling. Taken together, these results advance the application of graphene derivatives grafted with the biomolecules in the field of biosensing.