Honey Bee Allergen Api m3 Cloning for Desensitization
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Solution Overview
Problem
Current methods lack a reliable nucleic acid for recombinant production of acid phosphatase (Api m3) from honey bee venom, essential for desensitization therapy and diagnostic tests for insect venom allergies, due to incomplete cloning and conflicting data on the protein and nucleic acid sequences.
Innovation Solution
A nucleic acid encoding a polypeptide with a homology of more than 70% to the amino acid sequence of Api m3 is developed, allowing for successful amplification and verification of the full-length cDNA sequence, enabling recombinant production and diagnostic applications.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If incomplete cloning methods are used for Api m3, then the cloning process can be performed, but the sequence accuracy and reliability are insufficient
Solution Approach 1:
The patent divides the incomplete cloning process into multiple segments: initial cloning attempts, sequence verification, identification of conflicts, and iterative refinement. By segmenting the cloning process, the patent systematically addresses sequence accuracy issues while managing complexity through structured problem-solving steps.
Solution Approach 2:
The patent performs excessive verification actions beyond standard cloning protocols, including multiple sequence alignments, homology comparisons, and conflict resolution steps. This partial or excessive action ensures sequence accuracy by thoroughly validating each cloned sequence against known Api m3 characteristics and identifying discrepancies.
2Adaptability or versatility
If conflicting data on Api m3 sequences is used, then various studies can be conducted, but the diagnostic and therapeutic reliability is compromised
Solution Approach 1:
The patent introduces an intermediary verification process that mediates between conflicting sequence data from different studies. This intermediary layer performs homology analysis, identifies consistent regions, and resolves conflicts before final sequence determination, thereby maintaining research flexibility while ensuring diagnostic reliability.
Solution Approach 2:
The patent implements feedback mechanisms where cloned sequences are continuously verified against established Api m3 characteristics and previous study data. Discrepancies trigger iterative refinement cycles, allowing the research to adapt to new findings while maintaining reliability through consistent validation criteria.
3Productivity
If traditional cloning methods are used without full-length verification, then initial cloning can be achieved, but complete and accurate Api m3 sequence cannot be obtained
Solution Approach 1:
The patent performs preliminary actions by designing cloning strategies that anticipate completeness verification needs. Restriction sites are strategically selected, and cloning vectors are chosen to facilitate full-length sequence recovery, preventing incomplete cloning before it occurs while maintaining efficient productivity.
Solution Approach 2:
The patent replaces traditional mechanical cloning verification methods with molecular biology techniques such as restriction fragment analysis, sequencing, and homology comparison. This substitution enables comprehensive sequence verification without proportionally increasing cloning time, maintaining productivity while achieving complete sequence accuracy.
Data Source
AI summary
The present invention relates to a nucleic acid encoding a polypeptide capable of binding to IgE from subjects allergic to venom of an insect from the order Hymenoptera having a homology of more than 70% to the amino acid sequence of SEQ ID NO: 2, which is the honey bee allergen Api m3 (acid phosphatase). The invention further relates to expression vectors, host cells and polypeptides encoded by the nucleic acid, as well as diagnostic and pharmaceutical uses thereof.