The selection of highly specific and selective aptamers using modified SELEX and their use in process analytical techniques for Lucentis bioproduction

Bhardwaj, Tanu; Rathore, Anurag S.; Jha, Sandeep Kumar

doi:10.1039/d0ra03542d

Cited by 13 publications

(15 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The sequence of the ssDNA aptamer used for on-chip detection of Lucentis was (aptamer sequence 1 from our previous study on the selection of aptamers for Lucentis): 23 5′-AGGGAAGAGAAGGACATATGATGGGGGCTAGTATC-GACGTCTGCCTATGTTGTTAGTCGGTGTTGACTAGTACAT-GACCACTTG-3′.…”

Section: Methodsmentioning

confidence: 99%

“…In our previous study, we reported the selection of specific aptamers for Lucentis using Systematic evolution of ligands by exponential enrichment (SELEX). 23 Additionally, an aptamer biosensor using non-faradaic impedimetric spectroscopy (NFIS) was fabricated to detect Lucentis in the fermenter for proof-of-concept. Major changes in the capacitive component of impedance were noticed in the biosensor upon complex formation between aptamer and Lucentis.…”

mentioning

confidence: 99%

See 1 more Smart Citation

On-Line PAT Monitoring of Lucentis in the Fermenter Using Aptamer-Based Capacitive Microfluidic Chip Biosensor

Bhardwaj

Jha

2022

J. Electrochem. Soc.

Self Cite

View full text Add to dashboard Cite

In this study, we have developed an aptamer-based PDMS/glass microfluidic chip biosensor employing the capacitance based method of detection, which has the tendency to work as an on-line process analytical technology (PAT) monitoring tool to detect Lucentis in the fermenters. The limit of detection and linear range of detection of the chip was determined to be 8.5 and 8.5−100 nM, respectively, which was better than HPLC based method of Lucentis estimation. In real sample analysis, a significant correlation was obtained between the reported microchip based biosensor and HPLC-based detection method. Moreover, pre-concentration or pre-processing of media broth was not necessary with our method, which reduced the analysis time to 30 min. The chip was determined to be miniaturized, selective, specific, interference-free, label-free, rapid, and reusable with a long shelf-life. Additionally, the chip demonstrated the requirement of a small direct sample volume of 10 µl and simple automation using a peristaltic pump. The chip biosensor is expected to have on-line PAT monitoring use for checking the production of Lucentis in the fermenter leading to a reduction in the cost of the therapy and better affordability of the treatment to the age-related macular degeneration patients.

show abstract

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

On-Line PAT Monitoring of Lucentis in the Fermenter Using Aptamer-Based Capacitive Microfluidic Chip Biosensor

Bhardwaj

Jha

2022

J. Electrochem. Soc.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Agarose beads have also been used as SELEX platforms and used to select aptamers for diverse chemicals [64][65][66]69,86]. Agarose beads are linear hydrogel polymers comprising repeated units of agarobiose.…”

Section: Bead-mediated Aptamer Selectionmentioning

confidence: 99%

“…The aptamer for Lucentis was selected by immobilizing the target on the agarose beads [ 66 ]. An aptamer with high affinity was obtained by decreasing the concentration of the immobilized target and the incubation time, increasing the number of washing steps, as well as expanding the elution procedure, which could reduce the number of cycles.…”

Section: Current Aptamer Discovery Strategies To Detect Chemicals and Biomoleculesmentioning

confidence: 99%

Recent Advances in Micro/Nanomaterial-Based Aptamer Selection Strategies

Kim

Lee

et al. 2021

Molecules

View full text Add to dashboard Cite

Aptamers are artificial nucleic acid ligands that have been employed in various fundamental studies and applications, such as biological analyses, disease diagnostics, targeted therapeutics, and environmental pollutant detection. This review focuses on the recent advances in aptamer discovery strategies that have been used to detect various chemicals and biomolecules. Recent examples of the strategies discussed here are based on the classification of these micro/nanomaterial-mediated systematic evolution of ligands by exponential enrichment (SELEX) platforms into three categories: bead-mediated, carbon-based nanomaterial-mediated, and other nanoparticle-mediated strategies. In addition to describing the advantages and limitations of the aforementioned strategies, this review discusses potential strategies to develop high-performance aptamers.

show abstract

“…Aptamer sequences can bind with high specificity for their intended target, resulting in E-AB sensors with correspondingly high specificity and selectivity. [26][27][28][29][30] Why Focus on Electrochemical, Aptamer-Based Sensors?…”

mentioning

confidence: 99%

Perspective—Assessing Electrochemical, Aptamer-Based Sensors for Dynamic Monitoring of Cellular Signaling

Rousseau,

Kumakli,

White

2023

ECS Sens. Plus

View full text Add to dashboard Cite

Electrochemical, aptamer-based (E-AB) sensors provide a generalizable strategy to quantitatively detect a variety of targets including small molecules and proteins. The key signaling attributes of E-AB sensors (sensitivity, selectivity, specificity, and reagentless and dynamic sensing ability) make them well suited to monitor dynamic processes in complex environments. A key bioanalytical challenge that could benefit from the detection capabilities of E-AB sensors is that of cell signaling, which involves the release of molecular messengers into the extracellular space. Here, we provide a perspective on why E-AB sensors are suited for this measurement, sensor requirements, and pioneering examples of cellular signaling measurements.

show abstract

The selection of highly specific and selective aptamers using modified SELEX and their use in process analytical techniques for Lucentis bioproduction

Abstract: Highly specific and selective aptamers for Lucentis were selected using 10 rounds of a modified and highly stringent SELEX process.

Cited by 13 publications

References 27 publications

On-Line PAT Monitoring of Lucentis in the Fermenter Using Aptamer-Based Capacitive Microfluidic Chip Biosensor

On-Line PAT Monitoring of Lucentis in the Fermenter Using Aptamer-Based Capacitive Microfluidic Chip Biosensor

Recent Advances in Micro/Nanomaterial-Based Aptamer Selection Strategies

Perspective—Assessing Electrochemical, Aptamer-Based Sensors for Dynamic Monitoring of Cellular Signaling

Contact Info

Product

Resources

About