Monoclonal anti-idiotypic antibodies as functional internal images of enzyme active sites: production of a catalytic antibody with a cholinesterase activity.

Izadyar, Ladan; Friboulet, Alain; Remy, Marie Hélène; Roseto, A.; Thomas, Daniel

doi:10.1073/pnas.90.19.8876

Cited by 139 publications

(60 citation statements)

References 21 publications

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“…The proposal is supported further by the successful application in obtaining antibodies with cholinesterase-like (9) and ␤-lactamase activities (10). Specifically, mice immunized with a monoclonal IgG, AE-2, that displayed both high specificity to the active site of human erythrocyte acetylcholinesterase (AcChoE) and strong inhibition of the enzyme activity produced a monoclonal IgM (antiidiotype) with activity similar to that of the parental enzyme (9).…”

mentioning

confidence: 76%

“…We have demonstrated previously that the hydrolytic activity of 9A8 is characterized by a relaxation of specificity toward both substrates and inhibitors as compared with AcChoE (9). A specific feature of serine esterases, and of AcChoE in particular, is the specific irreversible inhibition by organophosphorous compounds (19).…”

Section: Resultsmentioning

confidence: 99%

“…To avoid possible contamination of mAb preparations by enzymes, efficient protocols for purification of the IgM abzyme were developed. It also has been shown that the parental AE-2 mAb inhibits the catalytic activity of 9A8 (9). To determine binding parameters of the idiotypic͞antiidiotypic mAb pair, we studied interaction of AE-2 with 9A8 by surface plasmon resonance (18) (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Enzyme mimicry by the antiidiotypic antibody approach

Kolesnikov

Kozyr

Alexandrova

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

110

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The concept of ''internal image'' of antiidiotypic antibodies has provided the basis for eliciting catalytic antibodies. A monoclonal IgM 9A8 that was obtained as an antiidiotype to AE-2 mAb, a known inhibitor of acetylcholinesterase, displayed esterolytic activity. Study of recombinant Fab fragments and separate light and heavy chains of 9A8 confirmed that the antibody variable domain encodes the catalytic function, whereas neither part of the primary sequence of the Fab exhibited homology with the enzyme. The specific modification of the 9A8 variable domain by an active site-directed covalent inhibitor revealed the presence of an active site Ser residue. A three-dimensional modeling suggests the existence of a functional catalytic dyad Ser-His. Comparison of active sites of 9A8 and 17E8 esterolytic abzyme raised against transitionstate analog revealed structural similarity although both antibodies were elicited by two different approaches.

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mentioning

confidence: 76%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Enzyme mimicry by the antiidiotypic antibody approach

Kolesnikov

Kozyr

Alexandrova

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

110

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“…Immunization of autoimmune-prone MRL-lpr/lpr mice with DNA-protein complexes also results in significantly higher production of anti-DNA Abs and abzymes with DNase activity comparing with nonautoimmune CBA and BALB/c healthy mice [29][30][31]. At the same time, artificial antiidiotypic abzymes can be induced by immunization of animals with different enzymes [23][24][25][26][27][28]. It was first suggested that natural SLE DNase Abzs may be antiidiotypic abzymes to topoisomerase I [53].…”

Section: Introductionmentioning

confidence: 97%

“…At the same time, some antiidiotypic Abs against active centers of many enzymes are also catalytically active [8,[12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28]. Healthy humans do more often not demonstrate catalytic Abs or their activities are extremely low.…”

Section: Introductionmentioning

confidence: 99%

Catalytic Antibodies in Norm and Systemic Lupus Erythematosus

Nevinsky¹

2017

Lupus

321

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Systemic lupus erythematosus (SLE) is known as a systemic polyethiologic diffuse autoimmune disease characterized by connective tissue disorganization and the paramount damage of skin and visceral capillaries. Usually, SLE symptoms include high fever, hair loss, mouth ulcers, chest pain, swollen lymph nodes, painful and swollen joints, increased fatigue, and appearance of red rash more often on the face. The exact reason of SLE appearance is not really clear. Detection of catalytic Abs (abzymes) was shown to be the earliest indicator of different AI disease development. Some abzymes are cytotoxic and can play a dangerous negative role in the pathogenesis of AI diseases. SLE is characterized by the appearance of abzymes with several different catalytic functions including hydrolysis of peptides and proteins, DNA, RNA, and oligosaccharides. In addition, monoclonal SLE abzymes are characterized by extraordinary diversity in the affinity to the substrates, physicochemical and catalytic characteristics, optimal conditions of catalysis, cytotoxicity, etc. Production of abzymes in SLE mice is associated with changes in the differentiation of hematopoietic stem cells of bone marrow, increase in lymphocyte proliferation, and significant suppression of cell apoptosis in different organs. In this chapter, abzymes with different catalytic activities in SLE are described.

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Detection of unwanted protein-bound ligands by capillary zone electrophoresis: The case of hidden ligands that stabilize cholinesterase conformation

2002

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Detection, identification and characterization of compounds present in purified proteins and biopharmaceuticals are of central interest. As well as chemical remedies, proteins of pharmacological interest have to exhibit their nakedness to become therapeutic drugs. Cholinesterases (ChE) are enzymes of major importance for detoxification of poisonous esters. Likewise, ChE are characterized by the high catalytic efficiency of an active site positioned at the bottom of a deep gorge. The gorge can be partially or fully occupied by ligands, i.e., substrates and inhibitors that are currently used in affinity chromatography purification steps. Accordingly, a suitable method allowing to analyse the presence of unwanted ligands and its influence on the functional conformation and stability of these enzymes was essential. We have developed CZE approaches for that purpose. The factors causing discrepancies between data for thermal unfolding of ChE by electrophoretic and by calorimetric methods were investigated. The presence of unwanted hidden ligands bound to purified enzymes was first demonstrated. The incidence of these ligands was discussed. Altogether, our results raised several questions concerning the real conformation of the native state of enzymes. Finally, CZE was proved to be a pertinent tool to validate the conformity of purified enzymes to a status of biopharmaceutical.

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Monoclonal anti-idiotypic antibodies as functional internal images of enzyme active sites: production of a catalytic antibody with a cholinesterase activity.

Abstract: Monodonal antibody 9A8 was selected by immunizing mice with AE-2, a monodonal antibody directed against the active ite of acetylchoilnesterase.

Cited by 139 publications

References 21 publications

Enzyme mimicry by the antiidiotypic antibody approach

Enzyme mimicry by the antiidiotypic antibody approach

Catalytic Antibodies in Norm and Systemic Lupus Erythematosus

Detection of unwanted protein-bound ligands by capillary zone electrophoresis: The case of hidden ligands that stabilize cholinesterase conformation

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