Production of highly potent horse antivenom against the Thai cobra (Naja kaouthia)

Pratanaphon, Ronachai; Akesowan, Surasak; Khow, Orawan; Sriprapat, Supod; Ratanabanangkoon, Kavi

doi:10.1016/s0264-410x(97)00098-4

Cited by 43 publications

(39 citation statements)

References 11 publications

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“…2A). Others have also observed a rapid rise in IgG titres following venom-immunzation in camelids and horses [8], [17], [33], [34]. Thereafter, we observed an abrupt decrease in the humoral response after the second boost (Day 42, total of 3 injections).…”

Section: Discussionsupporting

confidence: 71%

“…Cobra venoms, and especially that of N. kaouthia , are believed to be weakly immunogenic because of the low molecular weight of their neurotoxins [19], [33], [34]. Therefore, we used, with our own modifications, a ‘low dose, low volume, multi-site’ venom-immunization protocol that has been shown to be effective at raising high IgG titres against cobra venoms in horses [33] and recently, in camels [17].…”

Section: Methodsmentioning

confidence: 99%

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In Vivo Neutralization of α-Cobratoxin with High-Affinity Llama Single-Domain Antibodies (VHHs) and a VHH-Fc Antibody

et al. 2013

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Small recombinant antibody fragments (e.g. scFvs and VHHs), which are highly tissue permeable, are being investigated for antivenom production as conventional antivenoms consisting of IgG or F(ab’)2 antibody fragments do not effectively neutralize venom toxins located in deep tissues. However, antivenoms composed entirely of small antibody fragments may have poor therapeutic efficacy due to their short serum half-lives. To increase serum persistence and maintain tissue penetration, we prepared low and high molecular mass antivenom antibodies. Four llama VHHs were isolated from an immune VHH-displayed phage library and were shown to have high affinity, in the low nM range, for α-cobratoxin (α–Cbtx), the most lethal component of Naja kaouthia venom. Subsequently, our highest affinity VHH (C2) was fused to a human Fc fragment to create a VHH2-Fc antibody that would offer prolonged serum persistence. After in planta (Nicotiana benthamiana) expression and purification, we show that our VHH2-Fc antibody retained high affinity binding to α–Cbtx. Mouse α–Cbtx challenge studies showed that our highest affinity VHHs (C2 and C20) and the VHH2-Fc antibody effectively neutralized lethality induced by α–Cbtx at an antibody:toxin molar ratio as low as ca. 0.75×:1. Further research towards the development of an antivenom therapeutic involving these anti-α-Cbtx VHHs and VHH2-Fc antibody molecules should involve testing them as a combination, to determine whether they maintain tissue penetration capability and low immunogenicity, and whether they exhibit improved serum persistence and therapeutic efficacy.

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Section: Discussionsupporting

confidence: 71%

Section: Methodsmentioning

confidence: 99%

In Vivo Neutralization of α-Cobratoxin with High-Affinity Llama Single-Domain Antibodies (VHHs) and a VHH-Fc Antibody

et al. 2013

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“…For example, Christensen used about 150 mg and 75 mg of each venom for mAV and pAV productions, respectively (Christensen, 1979). With a novel immunization protocol (Pratanaphon et al, 1997) the amounts of each venom used for mAV and pAV productions are comparable (Chotwiwatthanakun et al, 2001).…”

Section: The Amount Of Venom Used In the Immunizationmentioning

confidence: 99%

“…Depending on the adjuvant used, some horses immunized with a snake venom may not respond or may fail to yield a high enough antibody titer (Pratanaphon et al, 1997). For mAV production, these non-responders can either be discarded or used in immunization with other more immunogenic venoms.…”

Section: Biological Variations In Mav and Pav Productionsmentioning

confidence: 99%

Merit and Demerit of Polyvalent Snake Antivenoms

Ratanabanangkoon

2003

Journal of Toxicology: Toxin Reviews

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Polyvalent antivenoms contain specific antibodies capable of neutralizing a number of homologous venoms from different species/genera. They can save lives of victims of snake envenomations, even when the culprit snake has not been identified (the usual case, about 80% of the time), and a monovalent antivenom can not be chosen. They are useful in areas where there are too many poisonous species to produce monovalent antivenoms against all of them. It is now possible to prepare polyvalent antivenoms with high potencies comparable to those of the corresponding monovalent antivenoms. With good manufacturing processes, these antivenoms have been shown to cause few and minor adverse reactions. In addition, polyvalent antivenoms exhibit a wider range of paraspecific neutralization of venoms from different species/genera, even from distant geographic areas. Lastly, it is less expensive and easier to produce and handle a few polyvalent antivenoms than batteries of monovalent antivenoms.

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“…Adjuvants can also enhance efficacy and allow a lower dose and thereby increase the potency of antivenins (Pratanaphon et al, 1997) and reduce vaccine costs (Singh and O'Hagan, 1999). The mechanism of action for Freund´s incomplete adjuvant (FIA) is the promotion of the formation of depots of antigen at a site of immunization (Guy, 2007).…”

Section: Introductionmentioning

confidence: 99%

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Bondarenko¹,

Podhorec²,

Svinger³

et al. 2015

Turk. J. Fish. Aquat. Sci.

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Various induction protocols were compared with the aim to reach ovulation in northern pike (Esox lucius L.). The effectiveness of salmon D-Arg 6 Pro 9 NEt-GnRHa at two doses in combination with various additives were tested: 50 and 100 µg kg -1 (groups GnRHa 1 and 2), GnRHa at 50 and 100 µg kg -1 in combination with 8 mg kg -1 metoclopramide (groups 3 and 4), GnRHa at 50 and 100 µg kg -1 emulsified in Freund´s incomplete adjuvant (FIA) (groups 5 and 6), and GnRHa at 50 and 100 µg kg-1 emulsified with FIA 8 mg kg -1 and metoclopramide (groups GnRHa 7 and 8). These treatments were compared to indoor controlled conditions (Group ICC, injected with saline solution, without hormonal treatment) and two traditional treatments: 3 mg kg -1 carp pituitary (Group CP) and ambient outdoor conditions (Group AOC, injected with saline solution only, without hormonal treatment).All fish in Group CP and 70% of those in Group AOC ovulated. The latency periods for Group CP was 96±4 h and 264±58 h for Group AOC. One Group-3 female ovulated spontaneously 108 h post-injection, whereas none of the other GnRHa injected or Group ICC fish ovulated. The pseudogonadosomatic index was 19.3±5.9%; 18.5% and 17.8±7.5% in the AOC, Group 3 and CP, respectively. The fertilization rate reached 88.6±4.5%; 85.5±12.0% and 66.0±13.7% in AOC, Group 3 and CP, respectively. The hatching rate was 68.6±9.9%; 65.5±7.5% and 54.4±8.0% in AOC, Group 3 and CP, respectively.The ovarian fluid pH was significantly higher (8.27±0.03) in AOC compared to CP group (8.11±0.02) (P>0.05). Ovarian fluid of the spontaneously ovulated Group-3 fish had a pH of 8.35. There were no significant differences in egg size and weight among successfully reproduced groups -AOC and CP (P>0.05).

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Production of highly potent horse antivenom against the Thai cobra (Naja kaouthia)

Cited by 43 publications

References 11 publications

In Vivo Neutralization of α-Cobratoxin with High-Affinity Llama Single-Domain Antibodies (VHHs) and a VHH-Fc Antibody

In Vivo Neutralization of α-Cobratoxin with High-Affinity Llama Single-Domain Antibodies (VHHs) and a VHH-Fc Antibody

Merit and Demerit of Polyvalent Snake Antivenoms

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