Patatin-01 Antibody

What is patatin and why is it important in immunological research?

Patatin is the predominant storage protein in potato tubers, making up about 60% of total protein content. It exists as dimers of 40-42 kDa subunits without disulfide bridges . In immunological research, patatin (also known as Sol t 1) is significant as it has been characterized as the primary allergen in potatoes. Studies have demonstrated the presence of IgE antibodies to Sol t 1 in individuals with potato allergies, particularly in atopic dermatitis patients .

The importance of patatin in immunological research extends beyond potato biology to serve as a model system for:

• Understanding plant food allergen properties and epitope structures

• Studying heat-stable food allergens (patatin maintains allergenicity after cooking)

• Investigating cross-reactivity patterns among plant allergens

• Developing improved diagnostic approaches for food allergies

How is Anti-Patatin Antibody used in potato protein research?

Anti-Patatin Antibody serves as a critical tool in potato protein research for multiple applications:

• Protein quantification: Detecting and measuring patatin levels in different potato varieties, tissues, or under various growth conditions

• Protein localization: Determining the spatial distribution of patatin within potato tuber cells and tissues through immunolocalization techniques

• Protein isolation: Selectively extracting patatin from complex potato protein mixtures using immunoprecipitation

• Western blotting: Identifying patatin in protein samples separated by electrophoresis, confirming protein identity and integrity

• Tracking protein changes: Monitoring patatin expression during potato development, storage, or in response to environmental stresses

These applications provide valuable insights into potato biology, protein storage mechanisms, and potential agricultural improvements.

What is the relationship between patatin and potato allergies?

Patatin (Sol t 1) has been established as the primary allergen in potatoes. Research has revealed several important aspects of this relationship:

• IgE reactivity: Studies show that 75% of infants suspected of having potato allergy had IgE antibodies to Sol t 1 (patatin) in enzyme-linked immunosorbent assay (ELISA)

• Heat stability: Unlike many food allergens that lose allergenicity upon cooking, patatin demonstrates heat stability. Research found positive skin-prick tests to natural Sol t 1 in 50% of potato-allergic infants, with challenge responses to both raw and cooked potato

• Clinical manifestations: Reactions to patatin can present as immediate hypersensitivity or delayed reactions like exacerbation of atopic dermatitis (AD). In one study, oral challenge with cooked potato resulted in one infant showing an immediate reaction while seven exhibited delayed reactions with worsening of AD

• Allergenic epitopes: Anti-patatin antibodies help identify the specific epitopes responsible for allergic reactions

This relationship makes patatin an important target for diagnostic testing in suspected potato allergies and a valuable model for studying food allergen properties.

How do researchers extract and purify patatin for antibody production?

Extracting and purifying patatin for antibody production involves several key methodological steps:

Table 1: Patatin Extraction and Purification Protocol

The purified patatin or patatin-derived peptides are then used to immunize host animals (commonly rabbits for polyclonal antibodies) for antibody production.

What applications are most suitable for Anti-Patatin Antibody use in laboratory settings?

Anti-Patatin Antibody demonstrates utility across multiple experimental applications, each with specific optimization requirements:

How does the specificity of Anti-Patatin Antibody differ when targeting various patatin isoforms?

The specificity of Anti-Patatin Antibody across different patatin isoforms is influenced by several factors:

• Isoform diversity: Potato patatin exists in multiple isoforms, with at least 36 known variants

• Epitope selection: Commercial antibodies often target conserved regions to detect multiple isoforms, such as those generated using "KLH-conjugated synthetic peptide derived from a C-terminal part of 36 known isoforms including Q3YJS9, Q3YJT0, Q42502, Q3YJT2"

• Cross-reactivity profiling methods:

  • Western blotting with purified isoforms

  • Competitive binding assays

  • Epitope mapping techniques

  • Pre-absorption controls with specific isoforms

• Experimental implications:

  • Broad-specificity antibodies detect total patatin content but may mask isoform-specific changes

  • Isoform-specific antibodies enable precise tracking of particular variants but may miss total patatin dynamics

• Validation approaches:

  • Knockout/knockdown controls using potato varieties lacking specific patatin isoforms

  • Recombinant protein controls with individually expressed patatin isoforms

  • Mass spectrometry correlation confirming antibody specificity through peptide identification

Researchers should select anti-patatin antibodies based on whether their experimental questions require pan-patatin detection or isoform-specific analysis.

What are the advantages and limitations of using polyclonal versus monoclonal antibodies for patatin detection?

Comparing polyclonal and monoclonal antibodies for patatin detection reveals distinct advantages and limitations:

Table 3: Polyclonal vs. Monoclonal Anti-Patatin Antibodies

Many researchers adopt a complementary approach, using both antibody types to leverage their respective strengths in patatin research.

What cross-reactivity concerns exist when using Anti-Patatin Antibody in complex biological samples?

When using Anti-Patatin Antibody in complex biological samples, researchers should address several cross-reactivity concerns:

• Patatin-like protein family cross-reactivity:

  • Mammalian samples may cross-react with patatin-like phospholipase domain-containing proteins (PNPLAs) such as:

    • PNPLA1 (Patatin-Like phospholipase Domain Containing 1)

    • PNPLA2/ATGL (Patatin Like Phospholipase Domain Containing Protein 2)

  • Microbial samples may cross-react with pathogen phospholipases:

    • TgPL3 from Toxoplasma gondii

    • Pat1 from Rickettsia parkeri

• Other plant protein cross-reactivity:

  • Related Solanaceae species may contain patatin homologs (tomato, eggplant, peppers)

  • Structurally similar storage proteins from unrelated plant species

• Assessment and mitigation strategies:

  • Pre-absorption controls with purified potential cross-reactive proteins

  • Western blot profiling against extracts from various species

  • Knock-out/knock-down validation using samples lacking target protein

  • Epitope analysis via in silico comparison across related proteins

  • Competitive binding assays with labeled and unlabeled potential cross-reactants

• Application-specific manifestations:

  • Immunohistochemistry: Unexpected staining patterns in non-target tissues

  • Western blotting: Additional bands at unexpected molecular weights

  • Immunoprecipitation: Co-precipitation of cross-reactive proteins

  • ELISA: Falsely elevated quantification values

Addressing these cross-reactivity concerns is essential for generating reliable, interpretable data with Anti-Patatin Antibody.

How does patatin-like phospholipase activity influence experimental design when using these antibodies?

Patatin and patatin-like proteins possess phospholipase activity that significantly impacts experimental design:

• Functional enzymatic considerations:

  • Patatin exhibits phospholipase A2 activity, hydrolyzing membrane phospholipids

  • This catalytic function is shared across patatin-like phospholipases:

    • Pat1 in Rickettsia parkeri mediates membrane escape and cell-cell spread

    • TgPL3 in Toxoplasma gondii contributes to host cell invasion

    • Human PNPLAs involved in lipid metabolism

• Epitope accessibility impacts:

  • Antibodies targeting the catalytic site may:

    • Be inhibited by substrate binding

    • Inhibit enzymatic activity themselves

    • Show differential binding to active versus inactive conformations

  • Substrate binding may induce structural changes affecting epitope exposure

• Sample preparation considerations:

  • For preserving enzymatic activity:

    • Avoid detergents that disrupt native structure

    • Consider native gel electrophoresis for Western blotting

    • Use mild fixation for immunohistochemistry

  • For inhibiting activity when it interferes with detection:

    • Include specific phospholipase inhibitors during sample preparation

    • Completely denature samples before antibody application

• Experimental controls design:

  • Use site-directed mutagenesis to create enzymatically inactive proteins as controls

  • Include phospholipid substrates during antibody incubation to assess binding interference

  • Incorporate probes that report on protein conformation alongside antibody detection

Understanding the dual nature of these proteins as both structural entities and active enzymes enables more effective experimental design and more accurate interpretation of results.

What are the key structural features of patatin that antibodies target?

Patatin possesses several structural features that serve as important epitopes for antibody targeting:

Figure 1: Major Structural Features of Patatin Targeted by Antibodies

• Conserved domains:

  • Anti-patatin antibodies often target conserved regions shared among numerous patatin isoforms

  • Commercial antibodies frequently use "synthetic peptide derived from a C-terminal part of 36 known isoforms of patatin from Solanum tuberosum"

  • These include regions containing sequences from variants Q3YJS9, Q3YJT0, Q42502, and Q3YJT2

• Quaternary structure:

  • Mature patatin exists as dimers of 40-42 kDa subunits without disulfide bridges

  • Some antibodies recognize epitopes formed at the dimer interface or exposed only in the dimeric state

  • Sample preparation conditions may affect these conformational epitopes

• Catalytic domain:

  • Patatin contains a catalytic domain with phospholipase A2 activity

  • This domain shares structural similarity with mammalian phospholipases, forming the basis for the "patatin-like phospholipase" protein family

  • Antibodies may target unique features of this catalytic region

• Surface-exposed regions vs. internal epitopes:

  • For applications like immunolocalization, antibodies typically target surface-exposed epitopes

  • For Western blotting, antibodies recognizing internal (linear) epitopes may be preferred

  • Application versatility often requires targeting epitopes that remain accessible under various conditions

Understanding these structural features helps researchers select appropriate antibodies for specific experimental applications and correctly interpret results.

What are the challenges in developing highly specific antibodies against patatin?

Developing highly specific antibodies against patatin presents several technical challenges:

• Isoform complexity:

  • The existence of at least 36 known patatin isoforms complicates antibody development

  • Creating pan-specific antibodies requires identifying perfectly conserved epitopes

  • Developing isoform-specific antibodies demands identifying unique regions in highly similar sequences

• Structural considerations:

  • Native patatin's dimeric structure presents conformational epitopes that may be lost in denatured conditions

  • The protein's three-dimensional folding can shield potential epitopes, limiting accessibility

  • Maintaining consistent protein conformation during immunization is challenging

• Post-translational modifications:

  • Variations in glycosylation or other modifications across patatin isoforms affect recognition

  • Antibodies may show different affinities for modified versus unmodified forms

  • Recombinant expression systems may not reproduce native modifications

• Cross-reactivity management:

  • Patatin's membership in the broader patatin-like phospholipase family creates specificity challenges

  • Preventing cross-reactivity with human patatin-like phospholipases (PNPLA1, PNPLA2) requires careful epitope selection

  • Balancing specificity with sufficient affinity presents optimization challenges

• Application versatility constraints:

  • Developing antibodies that perform consistently across multiple applications is difficult

  • Epitopes suitable for Western blotting may not be ideal for immunoprecipitation or immunohistochemistry

  • Optimizing for multiple applications often requires compromise

Addressing these challenges typically involves comprehensive epitope analysis, multiple immunization strategies, and extensive validation across applications and conditions.

How do researchers validate Anti-Patatin Antibody performance across different experimental contexts?

Rigorous validation of Anti-Patatin Antibody across experimental contexts involves a multi-faceted approach:

Table 4: Comprehensive Antibody Validation Framework

This comprehensive validation approach ensures reliable, reproducible results and helps researchers interpret findings accurately.

How can Anti-Patatin Antibody research contribute to allergy diagnostics and treatment?

Recent advances in using Anti-Patatin Antibody for allergenicity research have expanded our understanding of food allergies and improved diagnostic approaches:

• Heat-stable allergen characterization:

  • Patatin (Sol t 1) has been identified as a heat-stable potato allergen, maintaining allergenicity after cooking

  • Studies show IgE antibodies and positive skin-prick tests to Sol t 1 in potato-allergic infants

  • This research clarifies why cooked potato can still trigger allergic reactions in sensitive individuals

• Improved diagnostic approaches:

  • Component-resolved diagnostics using purified patatin (detected with anti-patatin antibodies) provide more precise allergy diagnosis than whole potato extracts

  • Standardized immunoassays using anti-patatin antibodies allow consistent quantification of potato allergens in food products

  • Anti-patatin antibodies enable development of sensitive assays for detecting potato allergens in processed foods

• Allergen epitope mapping:

  • Anti-patatin antibodies help identify immunodominant epitopes for targeted immunotherapy

  • Competitive binding studies between patient IgE and anti-patatin antibodies map allergenic epitopes

  • This research contributes to developing more effective allergy treatments

• Cross-reactivity characterization:

  • Antibodies targeting conserved epitopes in patatin help identify cross-reactive proteins in other foods

  • This approach helps predict cross-reactivity patterns and informs allergen avoidance recommendations

  • Understanding molecular patterns of cross-reactivity improves management of multiple food allergies

• Exposure assessment applications:

  • Beyond food, anti-patatin antibodies detect aerosolized potato proteins in occupational settings

  • This enables assessment of inhalation exposure risks and informs workplace safety measures

  • Similar approaches apply to other occupational allergens

These advances demonstrate how anti-patatin antibodies contribute to both fundamental research and practical applications in managing plant food allergies.

How do patatin and patatin-like proteins compare across species and research contexts?

Recent technological advances are transforming how researchers develop and apply anti-patatin antibodies:

• Antibody engineering approaches:

  • Phage display libraries enable rapid screening of antibody variants with enhanced specificity for patatin epitopes

  • Single-amino acid modifications can dramatically improve antibody performance, as demonstrated in related antibody development work

  • Computational antibody design methods combine deep learning with linear programming to optimize antibody properties

• Advanced detection systems:

  • Super-resolution microscopy enables nanoscale visualization of patatin distribution in cellular compartments

  • Multiplex immunoassays allow simultaneous detection of multiple patatin isoforms or patatin alongside other proteins

  • Single-molecule detection methods increase sensitivity for trace amounts of patatin

• Mass spectrometry integration:

  • Immuno-mass spectrometry combines antibody enrichment with MS analysis for enhanced specificity

  • Targeted mass spectrometry approaches like PRISM (high-pressure, high-resolution separations with intelligent selection and multiplexing) complement antibody-based methods

  • These integrative approaches validate antibody specificity while providing additional structural information

• High-throughput validation platforms:

  • Protein microarrays enable testing antibody specificity against thousands of potential cross-reactants

  • Automated liquid handling systems standardize validation protocols across laboratories

  • These platforms generate comprehensive validation datasets that enhance antibody reliability

• Novel application development:

  • Biosensor integration with anti-patatin antibodies creates sensitive food allergen detection systems

  • Antibody-based imaging probes enable in vivo tracking of allergic responses

  • Point-of-care diagnostic devices bring laboratory-quality detection to field settings

These technological advances are improving antibody specificity, sensitivity, and application versatility while enabling more comprehensive validation—all critical factors for advancing patatin research.