Recombinant Sphingomyelinase D Antigen Matrix for Spider Venom Neutralization
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Solution Overview
Problem
Current treatments for violin spider bites, particularly those from Loxosceles genus, are inadequate due to the need for large quantities of venom for antivenom production and the presence of irrelevant proteins in horse-derived antivenoms, leading to adverse immune reactions and inefficacy in neutralizing the venom, as well as the difficulty in accurately diagnosing the species causing the bite.
Innovation Solution
Development of recombinant sphingomyelinase D proteins from Loxosceles species, such as L. boneti, L. reclusa, and L. laeta, which are used as immunogens to generate neutralizing antibodies and as components of an antigenic matrix for immunoaffinity purification of antivenoms, and a diagnostic system to determine the species of the spider causing the bite.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If horse-derived antivenoms are used to treat violin spider bites, then neutralization of venom is achieved, but adverse immune reactions occur due to irrelevant proteins
Solution Approach 1:
The patent extracts and isolates the specific neutralizing component (sphingomyelinase D) from the complex horse-derived antivenom mixture. By using recombinant sphingomyelinase D as a pure immunogen, the invention removes irrelevant proteins that cause adverse reactions while retaining the essential neutralizing activity against violin spider venom
Solution Approach 2:
The patent creates a recombinant copy of the sphingomyelinase D protein using molecular biology techniques. This recombinant protein serves as a simplified model that replicates the neutralizing function of the natural antivenom component without the harmful irrelevant proteins present in horse-derived preparations
2Quantity of substance
If large quantities of venom are used for antivenom production, then sufficient antivenom is produced, but the complexity and cost of venom collection and purification increases
Solution Approach 1:
The patent extracts the essential immunogenic component (sphingomyelinase D) from the complex venom mixture. By using this purified recombinant protein as immunogen, the invention eliminates the need to collect and process large quantities of whole venom, significantly simplifying the production process while maintaining adequate antivenom yield
Solution Approach 2:
The recombinant sphingomyelinase D serves as a synthetic copy that can be produced in large quantities through bacterial expression systems, replacing the need for complex venom collection from thousands of spiders and simplifying the production scale-up process
3Adaptability or versatility
If whole venom is used as immunogen, then broad coverage against multiple venom components is achieved, but specificity for the main toxic component is reduced
Solution Approach 1:
The patent extracts the specific sphingomyelinase D component from the whole venom mixture to create a purified recombinant immunogen. This extraction approach increases the specificity of the immune response toward the main toxic component while eliminating cross-reactivity with less important venom proteins
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The recombinant proteins effectively generate neutralizing antibodies against the venom, reducing the risk of adverse reactions and improving treatment efficacy, while the diagnostic system enables timely and accurate identification of the spider species, facilitating appropriate treatment.
Implementation Method 1
This enzyme binds to the cell membranes (epithelial, endothelial of the vascular tissue and red blood cells) hydrolyzing sphingolipids to subsequently release phosphoceramide and choline
Implementation Method 2
an antigenic matrix capable of specifically binding neutralizing antibodies of the arachnid's venom, useful for the purification by immunoaffinity of such sero-therapic and fab-therapic agents
Data Source
AI summary
The invention relates to the isolation, characterization and expression of DNA fragments encoding sphingomyelinases D from three species of Loxosceles genus spiders, namely L. boneti, L reclusa and L. laeta, and the toxoids thereof. The invention also relates to the production of active sphingomyelinases D and the toxoids thereof using recombinant means and to the use of same as an immunogen for the production in vertebrates of antibodies that neutralise the corresponding venom and the respective fragments F(ab′)2. The invention further relates to the use of recombinant sphingomyelinases D as part of an antigen matrix which can be used in the immunopurification of antibodies and the fragments thereof or as part of any diagnostic device used to obtain clinical confirmation that the causal agent of poisoning in a patient is a spider of the Loxosceles genus. In addition, the invention includes molecular vectors for the expression of the DNA fragments, strains comprising same, which can express Loxosceles sphingomyelinases D, and methods for the expression thereof.AAPorcentaje de identidad de aminoacidos entre necrotoxinasde diferentes especies de Loxosceles.especieBBLb1Lr1Lr2LaLiLl-H17Ll-H13Ll-1Ll-2Lb1—Lr191—Lr28490—La868887—Li82848880—Ll-H175760605959—Ll-H13576061586080—Ll-156595958599980—Ll-25760615959829482—AA . . . AMINO ACID PERCENTAGE OF IDENTITY BETWEEN NECROTOXINS OF DIFFERENT LOXOSCELES SPECIESBB . . . SPECIES