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Increasing the Resolution of Field-Flow Fractionation with Increasing Crossflow Gradients.
Larson, Nicholas R; Bou-Assaf, George M.
Afiliação
  • Larson NR; Pharmaceutical Operations & Technology, 225 Binney Street, Cambridge, Massachusetts 02142, United States.
  • Bou-Assaf GM; Pharmaceutical Operations & Technology, 225 Binney Street, Cambridge, Massachusetts 02142, United States.
Anal Chem ; 95(44): 16138-16143, 2023 11 07.
Article em En | MEDLINE | ID: mdl-37874938
ABSTRACT
The resolution of flow field-flow fractionation (flow FFF) depends primarily on the crossflow rate and its change over time. In this work, we demonstrate a method for modulation of the crossflow rate during separation that increases the peak-to-peak resolution of the resulting fractograms. In classical FFF methods, the crossflow rate is either maintained constant or decreased during the separation of the different species. In this work, higher resolution between peaks was achieved by a novel gradient method in which the crossflow is increased briefly during separation to allow stronger retention of the later eluting peaks. We first outline the theoretical basis by which improved separation is achieved. We confirm our hypothesis by quantifying the impact of increasing crossflow on the resolution between a monoclonal antibody monomer and its high-molecular-weight aggregate. We then demonstrate that this method is applicable to two different FFF methods (AF4 and HF5) and various pharmaceutically relevant samples (monoclonal antibodies and adeno-associated viruses). Finally, we hypothesize that increasing the force perpendicular to the laminar flow as described here is broadly applicable to all FFF methods and improves the quality of FFF-based separations.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Fracionamento por Campo e Fluxo Idioma: En Revista: Anal Chem Ano de publicação: 2023 Tipo de documento: Article País de afiliação: Estados Unidos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Fracionamento por Campo e Fluxo Idioma: En Revista: Anal Chem Ano de publicação: 2023 Tipo de documento: Article País de afiliação: Estados Unidos