Conjugation of Enzymes to Antibodies

Winston, Scott; Fuller, Steven A.; Evelegh, Michael J.; Hurrell, John G.R.

doi:10.1002/0471142727.mb1101s50

Cited by 12 publications

(7 citation statements)

References 4 publications

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“…The small portion of the anti-camel rIgG that recognizes nanobodies could be extracted using affinity chromatography on a special type of columns containing covalently immobilized nanobodies as capturing ligands. Furthermore, by a covalent conjugating of anti-camel rIgG to HRP or AP enzymes, the final step of immunoassays which requires conjugated secondary antibodies can be omitted [29].…”

Section: Discussionmentioning

confidence: 99%

Purification of polyclonal IgG specific for Camelid’s antibodies and their recombinant nanobodies

et al. 2016

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Camelid’ s heavy-chain antibody (HCAb) consists of only two heavy chains and lacks the two light chains together with the CH1 domain usually found in conventional immunoglobulins. A recombinant single antigen-binding entity, named VHH (or Nanobody®) was generated by reengineering the variable domains from HCAb. This study focuses on the detection of camelid´s immunoglobulins as well as their derivative nanobodies using a universal anti-camel antibody produced in rabbit (rIgG). Starting from a crude rabbit serum, a standard stock of rIgG (1 mg/ml) was prepared after purification by affinity chromatography using protein-A column. As expected, rIgG was able to detect camel antibodies in ELISA and immunoblotting, and its reactivity was equal against all different camel IgG subclasses, which were purified from serum by differential affinity chromatography on protein-G and -A. Interestingly, rIgG also recognized nanobodies since they were originally part of camel HCAbs, providing an alternative method to detect the corpus of these recombinant proteins rather than targeting their artificial tags. These data suggest that the anti-camel rIgG described here could be efficiently applied at different stages of nanobody technology, including the quantitation of the issued nanobodies and their detection when bound to target antigens.

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Section: Discussionmentioning

confidence: 99%

Purification of polyclonal IgG specific for Camelid’s antibodies and their recombinant nanobodies

et al. 2016

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“…This binding even is however uncontrollable but the studies have shown that the antibodies conjugated following this method are still applicable, i.e. they retain both the enzyme and the antibody activity [39][40][41]. During this binding the reactive HRP site remains unperturbed and this was observed by the unchanged spectrum of HRP (alone) when compared to antibody bound HRP.…”

Section: Resultsmentioning

confidence: 99%

Design and evaluation of an electrochemical immunosensor for measles serodiagnosis using measles-specific Immunoglobulin G antibodies

Mashazi¹,

Vilakazi²,

Nyokong³

2013

Talanta

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“…The cis-diol, in the presence of periodate, is easily oxidized to a dialdehyde, which subsequently is reacted with N-nucleophiles, such as amines or hydrazines to introduce a desired functionality. 30 In our hands, this procedure works very well, and we have prepared a number of RNAs modified with fluorescence dyes or with flavine derivatives. 31 Circularization strategies by 5 0 -3 0 -end joining In general, the preparation of circRNAs from linear precursors is very challenging, due to the negative entropy associated with the circularization step.…”

Section: Chain Assemblymentioning

confidence: 99%

In vitro circularization of RNA

Müller

Appel

2016

RNA Biology

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Over the past 2 decades, different types of circular RNAs have been discovered in all kingdoms of life, and apparently, those circular species are more abundant than previously thought. Apart from circRNAs in viroids and viruses, circular transcripts have been discovered in rodents more than 20 y ago and recently have been reported to be abundant in many organisms including humans. Their exact function remains still unknown, although one may expect extensive functional studies to follow the currently dominant research into identification and discovery of circRNA by sophisticated sequencing techniques and bioinformatics. Functional studies require models and as such methods for preparation of circRNA in vitro. Here, we will review current protocols for RNA circularization and discuss future prospects in the field.Abbreviations: AMP, adenosine monophosphate; AppRNA, 5 0 -adenylated RNA; ATP, adenosine triphosphate; bp, base pair; CEV-A, citrus exocortis viroid strain A; circRNA, circular RNA; ciRNA, circular intronic RNA; CuAAC, cupper catalyzed azide alkyne cycloaddition; EDC, ethyl-3-(3 0 -dimethylaminopropyl)-carbodiimide; HIV, human immunodeficiency virus; IEciRNA, intron-exon circular RNA; nt, nucleotide; PIE, permuted intron exon; T4 Dnl, T4 DNA Ligase; T4 Rnl, T4 RNA Ligase

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Conjugation of Enzymes to Antibodies

Cited by 12 publications

References 4 publications

Purification of polyclonal IgG specific for Camelid’s antibodies and their recombinant nanobodies

Purification of polyclonal IgG specific for Camelid’s antibodies and their recombinant nanobodies

Design and evaluation of an electrochemical immunosensor for measles serodiagnosis using measles-specific Immunoglobulin G antibodies

In vitro circularization of RNA

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