Antibody-receptor bioengineering and its implications in designing bioelectronic devices

Dkhar, Daphika S.; Kumari, Rohini; Mahapatra, Supratim; Divya, .; Kumar, Rahul; Tripathi, Timir; Chandra, Pranjal

doi:10.1016/j.ijbiomac.2022.07.109

Cited by 16 publications

(7 citation statements)

References 130 publications

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“…Consequently, the utilization of antibody fragments as bioreceptors, as opposed to full-length antibodies, can result in noticeable signal amplification. 46 The use of smaller-sized antibody fragments, such as Fab and scFv fragments, allows a higher density of target binding to be achieved in a given volume. 47 Antibody fragments can be produced by various genetic modifications or chemical reduction methods.…”

Section: Discussionmentioning

confidence: 99%

“…Previously, the generation of antibody fragments, specifically Fab and (Fab)2 0 , relied on proteolytic cleavage of classical antibodies. 46,48,49 However, Plückthun and Skerra have outlined an alternative approach using cloning vectors to produce fully functional Fv and Fab fragments. By employing cloning vectors, antibodies can be directed into the periplasmic space, benefitting from the oxidizing environment that facilitates disulfide bond formation.…”

Section: Discussionmentioning

confidence: 99%

“…In the realm of biosensing, the configurations created through antibody engineering can boost signals, given that the resistance posed by conventional antibodies significantly exceeds that of these fragments. Consequently, the utilization of antibody fragments as bioreceptors, as opposed to full‐length antibodies, can result in noticeable signal amplification 46 . The use of smaller‐sized antibody fragments, such as Fab and scFv fragments, allows a higher density of target binding to be achieved in a given volume 47 .…”

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Utilizing fab fragment‐conjugated surface plasmon resonance‐based biosensor for detection of SalmonellaEnteritidis

Eser,

Ekiz,

Ekiz

2024

J of Molecular Recognition

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Although antibodies, a key element of biorecognition, are frequently used as biosensor probes, the use of these large molecules can lead to adverse effects. Fab fragments can be reduced to allow proper antigen‐binding orientation via thiol groups containing Fab sites that can directly penetrate Au sites chemically. In this study, the ability of the surface plasmon resonance (SPR) sensor to detect Salmonella was studied. Tris(2‐carboxyethyl)phosphine was used as a reducing agent to obtain half antibody fragments. Sensor surface was immobilized with antibody, and bacteria suspensions were injected from low to high concentrations. Response units were changed by binding first reduced antibody fragments, then bacteria. The biosensor was able to determine the bacterial concentrations between 103 and 108 CFU/mL. Based on these results, the half antibody fragmentation method can be generalized for faster, label‐free, sensitive, and selective detection of other bacteria species.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Utilizing fab fragment‐conjugated surface plasmon resonance‐based biosensor for detection of SalmonellaEnteritidis

Eser,

Ekiz,

Ekiz

2024

J of Molecular Recognition

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“…The interest in developing new dedicated biosensors has expanded in parallel, both in academia and industry, attempting to exploit their potentiality in various new fields of application [1]. Among all biosensors, optical ones represent the most promising types which have been in most cases developed by exploiting the ability of surface-immobilized antibodies to recognize the target analytes with high affinity and specificity [2]. In such optical immune biosensors antibody immobilization is a crucial step, as it should ideally maintain the antigen recognition ability as in solution [3].…”

Section: Introductionmentioning

confidence: 99%

High sensitivity lab-on-fiber biosensing platform assisted by oriented antibody immobilization strategy

Ucci

Spaziani

Quero

et al. 2023

European Workshop on Optical Fibre Sensors (EWOFS 2023)

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The demand for highly sensitive, fast and low-cost biosensors for reliable quantification of small biomolecules or cancer biomarkers is leading to the development of a new class of devices able to change the techniques currently used for diagnosis in oncology. Lab‐on‐fiber (LoF) optrodes offer several advantages over conventional techniques for point‐of‐care platforms aimed at real‐time and label‐free detection of clinically relevant biomarkers. Moreover, the easy integration of LoF platforms in medical needles, catheters and nano-endoscopes offers unique potentials for in vivo biopsies and tumor microenvironment assessment. Here, we demonstrate the capability to improve the immobilization strategies through the use of hinge carbohydrates by involving homemade antibodies that demonstrated a significantly improved recognition of the antigen with ultra‐low detection limits. In order to create an effective pipeline for the improvement of biofunctionalization protocols to be used in connection with the LoF platform, here we first, optimized the protocol using a microfluidic Surface Plasmon Resonance device. Then we transferred the optimized strategy on LoF platform, based on Optical Fiber Meta‐tip (OFMT), for the final validation. As a clinically relevant scenario, we focused on a serological biomarker, Cripto‐1, for its ability to promote tumorigenesis in breast and liver cancer. Reported results demonstrate that the proposed approach based on oriented antibody immobilization is able to significantly improve Cripto‐1 detection with a ten‐fold enhancement versus the random approach. Therefore, our work opens new avenues in the development of high‐sensitivity LoF biosensors for the detection of clinically relevant biomarkers in the sub‐ng/mL range.

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“…The interest in developing new dedicated biosensors has expanded in parallel, both in academia and industry, while attempting to exploit their potentiality in various new fields of application [ 1 , 2 , 3 ]. Among all biosensors, optical ones represent the most promising types and in most cases have been developed to exploit the ability of surface-immobilized antibodies to recognize the target analytes with high affinity and specificity [ 4 , 5 ].…”

Section: Introductionmentioning

confidence: 99%

Advanced Lab-on-Fiber Optrodes Assisted by Oriented Antibody Immobilization Strategy

et al. 2022

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Lab-on-fiber (LoF) optrodes offer several advantages over conventional techniques for point-of-care platforms aimed at real-time and label-free detection of clinically relevant biomarkers. Moreover, the easy integration of LoF platforms in medical needles, catheters, and nano endoscopes offer unique potentials for in vivo biopsies and tumor microenvironment assessment. The main barrier to translating the vision close to reality is the need to further lower the final limit of detection of developed optrodes. For immune-biosensing purposes, the assay sensitivity significantly relies on the capability to correctly immobilize the capture antibody in terms of uniform coverage and correct orientation of the bioreceptor, especially when very low detection limits are requested as in the case of cancer diagnostics. Here, we investigated the possibility to improve the immobilization strategies through the use of hinge carbohydrates by involving homemade antibodies that demonstrated a significantly improved recognition of the antigen with ultra-low detection limits. In order to create an effective pipeline for the improvement of biofunctionalization protocols to be used in connection with LoF platforms, we first optimized the protocol using a microfluidic surface plasmon resonance (mSPR) device and then transferred the optimized strategy onto LoF platforms selected for the final validation. Here, we selected two different LoF platforms: a biolayer interferometry (BLI)-based device (commercially available) and a homemade advanced LoF biosensor based on optical fiber meta-tips (OFMTs). As a clinically relevant scenario, here we focused our attention on a promising serological biomarker, Cripto-1, for its ability to promote tumorigenesis in breast and liver cancer. Currently, Cripto-1 detection relies on laborious and time-consuming immunoassays. The reported results demonstrated that the proposed approach based on oriented antibody immobilization was able to significantly improve Cripto-1 detection with a 10-fold enhancement versus the random approach. More interestingly, by using the oriented antibody immobilization strategy, the OFMTs-based platform was able to reveal Cripto-1 at a concentration of 0.05 nM, exhibiting detection capabilities much higher (by a factor of 250) than those provided by the commercial LoF platform based on BLI and similar to the ones shown by the commercial and well-established bench-top mSPR Biacore 8K system. Therefore, our work opened new avenues into the development of high-sensitivity LoF biosensors for the detection of clinically relevant biomarkers in the sub-ng/mL range.

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Antibody-receptor bioengineering and its implications in designing bioelectronic devices

Cited by 16 publications

References 130 publications

Utilizing fab fragment‐conjugated surface plasmon resonance‐based biosensor for detection of SalmonellaEnteritidis

Utilizing fab fragment‐conjugated surface plasmon resonance‐based biosensor for detection of SalmonellaEnteritidis

High sensitivity lab-on-fiber biosensing platform assisted by oriented antibody immobilization strategy

Advanced Lab-on-Fiber Optrodes Assisted by Oriented Antibody Immobilization Strategy

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