Quantitative analysis of therapeutic proteins in biological fluids: recent advancement in analytical techniques

Song, Jae Geun; Baral, Kshitis Chandra; Kim, Gyu-Lin; Park, Ji Won; Seo, Soo-Hwa; Kim, Da-Hyun; Jung, Dong Hoon; Ifekpolugo, Nonye Linda; Han, Hyo-Kyung

doi:10.1080/10717544.2023.2183816

Cited by 6 publications

(5 citation statements)

References 201 publications

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“…The above proteins were prepared in buffer and stored at 4˚C for at least 24 hours before use. Protein concentrations were confirmed by UV-Vis at A 280 [4,31,32].…”

Section: Reagentsmentioning

confidence: 99%

“…Some of which were also examined previously [22]. PSV values given in Table 3 for HSA [20,23,31], lysozyme [2,4], SARS-CoV-2 RBD [48][49][50], ACE2 [48], streptavidin [51], and HSV-GpD [52] were taken from the literature. Table 3 shows for all cases examined, there was excellent agreement between protein concentrations estimated from the DNA standard curve (column 3) and concentrations determined by UV absorbance or measured dry weight (column 4).…”

Section: Plos Onementioning

confidence: 99%

“…Protein quantitation, the process of determining the exact concentration of a targeted protein in a sample, is an essential element of biological and chemical research, biotechnology development, drug discovery, and clinical diagnostic [1][2][3][4]. Specifically, accurate quantification of protein concentrations is required for determination of protein expression levels, monitoring protein-protein interactions, evaluating drug efficacy, identifying disease biomarkers, evaluating virus-host interactions, drug screening and development [1,2,4,5].…”

Section: Introductionmentioning

confidence: 99%

“…Classical techniques for determining concentrations of DNA and protein solutions include absorbance spectroscopy (UV-Vis), the bicinchoninic acid (BCA), Bradford, Lowry and fluorescence assays, the latter which have the added requirement of specific molecular probes [2,4,6,7]. UV-Vis, either from direct measurements or a colorimetric assay indicator, is the most widely employed method for quantitative determination of concentrations of nucleic acids and proteins in solution.…”

Section: Introductionmentioning

confidence: 99%

“…To determine accurate protein concentrations for a specific molecule, these methods typically require pure solutions. Even though standard methods can generally provide reliable measurements of protein concentrations, they can yield conflicting results [4,5,8]. For many cases UV-Vis measurements can provide accurate concentrations.…”

Section: Introductionmentioning

confidence: 99%

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DNA-based assay for calorimetric determination of protein concentrations in pure or mixed solutions

Eskew,

Reardon,

Benight

2024

PLoS ONE

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It was recently reported that values of the transition heat capacities, as measured by differential scanning calorimetry, for two globular proteins and a short DNA hairpin in NaCl buffer are essentially equivalent, at equal concentrations (mg/mL). To validate the broad applicability of this phenomenon, additional evidence for this equivalence is presented that reveals it does not depend on DNA sequence, buffer salt, or transition temperature (Tm). Based on the equivalence of transition heat capacities, a calorimetric method was devised to determine protein concentrations in pure and complex solutions. The scheme uses direct comparisons between the thermodynamic stability of a short DNA hairpin standard of known concentration, and thermodynamic stability of protein solutions of unknown concentrations. Sequences of two DNA hairpins were designed to confer a near 20°C difference in their Tm values. In all cases, evaluated protein concentrations determined from the DNA standard curves agreed with the UV-Vis concentration for monomeric proteins. For multimeric proteins evaluated concentrations were greater than determined by UV-Vis suggesting the calorimetric approach can also be an indicator of molecular stoichiometry.

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“…The above proteins were prepared in buffer and stored at 4˚C for at least 24 hours before use. Protein concentrations were confirmed by UV-Vis at A 280 [4,31,32].…”

Section: Reagentsmentioning

confidence: 99%