Utilization of Multi-Immunization and Multiple Selection Strategies for Isolation of Hapten-Specific Antibodies from Recombinant Antibody Phage Display Libraries

Tullila, Antti; Nevanen, Tarja K.

doi:10.3390/ijms18061169

Cited by 15 publications

(17 citation statements)

References 41 publications

(50 reference statements)

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“…In agreement with our previous results from the SPR assay using affinity in solution approach, the affinity constant for MPA and anti-MPA Fab antibody interaction was ∼40 nmol L –1 . The affinity of the interaction between cyclic peptide (A2) and anti-MPA Fab antibody is 2 orders of magnitude lower compared to the affinity of MPA–anti-MPA Fab antibody interaction.…”

Section: Resultssupporting

confidence: 91%

“…To compare the binding properties of the biotinylated cyclic peptide and MPA toward the anti-MPA antibody, label-free SPR technology was applied. In the binding experiments, previously identified, produced, and purified Fab antibodies recognizing either MPA or ochratoxin A were immobilized onto sensor chip surfaces . The same experimental conditions were used to study the binding properties of cyclic peptide (A2) and MPA.…”

Section: Resultsmentioning

confidence: 99%

“…NanoGlo Reagent for Immunoassay was from Promega Corporation (Madison, WI, USA), and High Capacity Magne Streptavidin Beads and ATG-42 plasmid DNA, containing the NanoLuc gene, were kindly donated by Promega Corporation (Madison, WI, USA). The recombinant anti-MPA Fab was obtained from a phage display library and produced as described previously …”

Section: Methodsmentioning

confidence: 99%

“…The recombinant anti-MPA Fab was obtained from a phage display library and produced as described previously. 22 Biopanning Rounds. A commercial phage-displayed peptide library was used to select cyclic peptides that bind to the anti-MPA.…”

Section: ■ Introductionmentioning

confidence: 99%

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Recombinant Peptide Mimetic NanoLuc Tracer for Sensitive Immunodetection of Mycophenolic Acid

et al. 2021

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Mycophenolic acid (MPA) is an immunosuppressant drug commonly used to prevent organ rejection in transplanted patients. MPA monitoring is of great interest due to its small therapeutic window. In this work, a phage-displayed peptide library was used to select cyclic peptides that bind to the MPA-specific recombinant antibody fragment (Fab) and mimic the behavior of MPA. After biopanning, several phage-displayed peptides were isolated and tested to confirm their epitope-mimicking nature in phage-based competitive immunoassays. After identifying the best MPA mimetic (ACEGLYAHWC with a disulfide constrained loop), several immunoassay approaches were tested, and a recombinant fusion protein containing the peptide sequence with a bioluminescent enzyme, NanoLuc, was developed. The recombinant fusion enabled its direct use as the tracer in competitive immunoassays without the need for secondary antibodies or further labeling. A bioluminescent sensor, using streptavidin-coupled magnetic beads for the immobilization of the biotinylated Fab antibody, enabled the detection of MPA with a detection limit of 0.26 ng mL –1 and an IC 50 of 2.9 ± 0.5 ng mL –1 . The biosensor showed good selectivity toward MPA and was applied to the analysis of the immunosuppressive drug in clinical samples, of both healthy and MPA-treated patients, followed by validation by liquid chromatography coupled to diode array detection.

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Section: Resultssupporting

confidence: 91%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

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Recombinant Peptide Mimetic NanoLuc Tracer for Sensitive Immunodetection of Mycophenolic Acid

et al. 2021

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“…However, the process is long and inherently dependent on animal immunization individually for each target. Phage-displayed scFv libraries have been constructed from immunized mice against FB 1 [64], OTA [65], and ZEN [66]. Surface plasmon resonance (SPR) analysis revealed excellent affinities in the nanomolar range for the anti-OTA and anti-ZEN antibody fragments, demonstrating the advantage of using immunized libraries.…”

Section: Recombinant Antibodiesmentioning

confidence: 99%

Bioinspired recognition elements for mycotoxin sensors

2017

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Mycotoxins are low molecular weight molecules produced as secondary metabolites by filamentous fungi that can be found as natural contaminants in many foods and feeds. These toxins have been shown to have adverse effects on both human and animal health, and are the cause of significant economic losses worldwide. Sensors for mycotoxin analysis have traditionally applied elements of biological origin for the selective recognition purposes. However, since the 1970s there has been an exponential growth in the use of genetically engineered or synthetic biomimetic recognition elements that allow some of the limitations associated with the use of natural receptors for the analyses of these toxins to be circumvented. This review provides an overview of recent advances in the application of bioinspired recognition elements, including recombinant antibodies, peptides, aptamers, and molecularly imprinted polymers, to the development of sensors for mycotoxins based on different transduction elements. Graphical abstract Novel analytical methods based on bioinspired recognition elements, such as recombinant antibodies, peptides, aptamers, and molecularly imprinted polymers, can improve the detection of mycotoxins and provide better tools than their natural counterparts to ensure food safety.

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Antibody‐Drug‐Conjugate Therapy for Hematological Cancers: Matching Cell Biology with Clinical Benefit

Firer

Luboshits

2021

Adv Funct Materials

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Progress in the development and clinical efficacy of antibody-drug conjugates (ADCs) has attracted the interests of both academic researchers and the industry. The rationale for developing ADCs is based on the inherent nonselectivity of chemotherapy for tumor cells, leading to side effects that can be severe and life threatening. Another major consequence of chemotherapy is the development of drug resistance. For these reasons, targeted drug delivery (TDD) systems are being developed to specifically deliver the cytotoxic drug into the target cell. A variety of carriers that can deliver drugs to the surface or into cancer cells, one of which is the use of monoclonal antibodies, with to date ten ADCs having received FDA approval. This review will focus on the development and clinical application of five ADCs currently approved for the treatment of hematological malignancies. Following a short history that led to their regulatory approval, the review will focus on the important link between the biology of the ADC, the cell surface component targeted by the antibody component, and clinical efficacy and conclude by highlighting newer developments that strengthen this link, so that ADCs can provide long-term clinical benefit to patients with hematological cancers.

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Utilization of Multi-Immunization and Multiple Selection Strategies for Isolation of Hapten-Specific Antibodies from Recombinant Antibody Phage Display Libraries

Cited by 15 publications

References 41 publications

Recombinant Peptide Mimetic NanoLuc Tracer for Sensitive Immunodetection of Mycophenolic Acid

Recombinant Peptide Mimetic NanoLuc Tracer for Sensitive Immunodetection of Mycophenolic Acid

Bioinspired recognition elements for mycotoxin sensors

Antibody‐Drug‐Conjugate Therapy for Hematological Cancers: Matching Cell Biology with Clinical Benefit

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