Resistance of antivenom proteins to foaming-induced denaturation.
Reconstitution of lyophilized antivenom is usually accomplished by gentle swirling for sometimes as much as 45min. Gentle resuspension is employed in order to avoid foaming the antivenom, which could have a variety of consequences including unfolding and denaturation of the protein, thus rendering it inactive. However, foaming of antivenom might cause only a small portion of the total protein to unfold or the protein may refold as foaming subsides. In this report, the effects of intentional severe foaming of three antivenom preparations are tested. Samples of gently resuspended antivenoms were subject to severe foaming by mechanical means, then tested for precipitation by examining the amounts of protein remaining in solution, for structural changes by circular dichroism spectroscopy and for activity changes by ELISA. In all cases, either no or minimal changes in antivenom properties were observed. These results indicate that severe intentional foaming does not substantially affect the properties of the antivenoms. It may be possible to employ more vigorous resuspension methods without affecting the efficacy of the antivenom. This could lead to a significant decrease in the time between envenomation and administration of antivenom.
Kanavage
Anthony D
AD
Department of Biochemistry and Molecular Biophysics, University of Arizona, BSW 362, P.O. Box 210088, Tucson, AZ 85721, USA.
Boyer
Leslie V
LV
McNally
Jude
J
Osterhout
John J
JJ
D000997:Antivenins / Q000737:chemistry*; D002942:Circular Dichroism / Q000379:methods*; D011489:Protein Denaturation; D017510:Protein Folding; D012910:Snake Venoms
D000997:Antivenins; D012910:Snake Venoms