Allergen immunotherapy (AIT) holds promise for inducing long-term immune tolerance and reducing symptoms, but it faces challenges such as lengthy treatment duration and potential side effects. To overcome the key limitations of traditional allergen immunotherapy (AIT), we integrated innovative strategies to enhance safety and efficacy. First, we rationally designed hypoallergenic derivatives of six major allergens by mutating IgE-binding epitopes while preserving T-cell reactivity, significantly reducing anaphylaxis risk. Second, we fused these hypoallergenic derivatives with the self-assembling peptide EFK8 to form broad-spectrum constructs, enabling multi-allergen targeting and extended half-life. Collectively, these approaches addressed AIT challenges related to side effects, complexity, and invasiveness while promoting robust immune tolerance.

In this project, the newly developed parts can be broadly categorized into two groups:

(1) Derivatives based on hypoallergenic variants of natural allergens.

(2) Fusion allergen constructs designed for enhanced AIT.

All parts have been successfully synthesized, expressed, and functionally validated.

Table 1 Part contribution about hypoallergenic derivatives

Part number	Part name	Part type	Contribution
BBa_25Z18GO2	mPhl p1	Basic part	New part
BBa_25YASPXX	mBet v1	Basic part	New part
BBa_25OYPV7W	mDer p1	Basic part	New part
BBa_25I9KJN2	mDer p2	Basic part	New part
BBa_258C9QZG	mMet e1	Basic part	New part
BBa_25BXZ798	mCyp c1	Basic part	New part
BBa_256CPPF4	pET32a-mPhl p1-His tag	Composite part	New part
BBa_25O5OJ6E	pET32a-mBet v1-His tag	Composite part	New part
BBa_25FIL2MS	pET32a-mDer p1-His tag	Composite part	New part
BBa_25VCKEJD	pET32a-mDer p2-His tag	Composite part	New part
BBa_25MPK9V8	pET32a-mMet e1-His tag	Composite part	New part
BBa_25380N0E	pET32a-mCyp c1-His tag	Composite part	New part

Derived from Phleum pratense (timothy grass pollen allergen). The sequence was synthetically constructed and reengineered to produce a hypoallergenic variant. The natural Phl p 1 sequence was segmented into four fragments and reassembled into a mosaic protein to disrupt conformational IgE-binding epitopes.

Properties:

mPhl p1 is a hypoallergenic recombinant allergen, engineered to reduce IgE-binding while retaining T-cell reactivity. This property allows it to induce immunological tolerance, making it a promising candidate for allergen-specific immunotherapy (AIT). It contains no native IgE epitopes, significantly lowering the risk of anaphylactic reactions.

Usage and Biology:

This part is used in allergen-specific immunotherapy (AIT) as a safer alternative to natural allergens. The recombinant hypoallergenic protein can stimulate regulatory T-cell responses and promote the production of allergen-specific IgG antibodies, which can competitively inhibit IgE binding. mPhl p1 targets Phl p 1-sensitized patients with allergic rhinitis or asthma due to grass pollen exposure. Studies have shown such engineered allergens can maintain immunogenicity while minimizing allergenicity (Ball et al., 2009).

Reference:

Ball, T., Linhart, B., Sonneck, K., Blatt, K., Herrmann, H., Valent, P., . . . Valenta, R. (2009). Reducing allergenicity by altering allergen fold: a mosaic protein of Phl p 1 for allergy vaccination. 64(4), 569-580. doi:https://doi.org/10.1111/j.1398-9995.2008.01910.x

A hypoallergenic variant of Bet v 1 with disrupted IgE-binding epitopes through 7 consecutive amino acid replacements, designed to reduce allergic reactions while retaining T-cell reactivity (Wallner et al., 2011).

Usage and Biology:

The mBet v1 protein is a hypoallergenic derivative of the major birch pollen allergen Bet v 1. It is engineered for allergen-specific immunotherapy (AIT) to treat birch pollen allergies. By mutating IgE-binding epitopes, mBet v1 minimizes the risk of anaphylactic reactions while preserving T-cell epitopes necessary for inducing immune tolerance. This protein can stimulate the production of protective IgG antibodies that block IgE binding, promoting long-term desensitization. Hypoallergenic variants like mBet v1 are critical for safer and more effective AIT, particularly for patients with severe allergies.

Reference:

Wallner, M., Hauser, M., Himly, M., Zaborsky, N., Mutschlechner, S., Harrer, A., . . . Ferreira, F. (2011). Reshaping the Bet v 1 fold modulates TH polarization. Journal of Allergy and Clinical Immunology, 127(6), 1571-1578.e1579. doi:https://doi.org/10.1016/j.jaci.2011.01.064

Gene synthesized based on the sequence of the Dermatophagoides pteronyssinus Der p1 allergen, containing five linked T cell epitopes (T1–T5). The gene was inserted into pET-28a(+) vector and expressed in E. coli BL21 (DE3).

●Properties:

mDer p1 T encodes a T-cell epitope fusion peptide derived from the major house dust mite allergen Der p1. It eliminates B-cell epitopes to reduce IgE binding, while preserving T-cell activatio, making it suitable for allergen-specific immunotherapy (SIT) with low allergenicity.

●Usage and Biology：

The Der p1 fusion peptide is a hypoallergenic derivative designed to address limitations of traditional allergen immunotherapy (AIT), such as standardization difficulties and systemic allergic side effects. By retaining only key T-cell epitopes and removing IgE-binding regions, this recombinant allergen can induce immune tolerance without triggering hypersensitivity.

Studies in asthmatic mice demonstrated that mDer p1 significantly reduced airway inflammation, serum IgE levels, and Th2/Th17 cytokines, while promoting Th1/Treg responses. These findings indicate that mDer p1 may be a promising candidate for safer, more effective AIT.

Reference:

Hong, Y. (2018). Exploration of the mechanism of Der p1 T-cell epitope fusion peptide-specific immunotherapy in house dust mite-sensitized asthmatic mice., Available from Cnki

A hypoallergenic variant of Der p 2 engineered through site-directed alanine mutagenesis at key IgE-binding residues (e.g., K96, E102), significantly reducing IgE reactivity while maintaining structural stability and T-cell epitope integrity. Mutant K96A exhibits near-complete loss of IgE binding due to misfolding, whereas E102A retains native folding with markedly decreased allergenicity.

Usage and Biology​​:

The hypoallergenic Der p 2 mutants are designed for safer allergen-specific immunotherapy (AIT) against dust mite allergy. By disrupting conformational IgE epitopes, these derivatives minimize anaphylaxis risk while preserving T-cell reactivity essential for immune tolerance induction. In murine models, immunization with these mutants elicits IgG antibodies that competitively inhibit human IgE binding to wild-type Der p 2. Additionally, they stimulate peripheral blood mononuclear cell (PBMC) proliferation comparable to the native allergen, confirming retained immunogenicity. These properties make them promising candidates for next-generation AIT vaccines aimed at achieving long-term desensitization with enhanced safety profiles.

Reference:

Reginald, K., & Chew, F. T. (2018). Conformational IgE Epitope Mapping of Der p 2 and the Evaluations of Two Candidate Hypoallergens for Immunotherapy. Scientific Reports, 8(1), 3391. doi:10.1038/s41598-018-21792-1

A truncated derivative generated by deleting all nine identified IgE-binding epitopes. Both variants were designed to significantly reduce or eliminate IgE reactivity while preserving structural integrity and T-cell epitopes essential for immunogenicity. 

​​Usage and Biology:​​

The engineered hypoallergen was designed for safer and more effective ​​allergen-specific immunotherapy (AIT)​​ against shrimp tropomyosin allergy. It exhibited markedly reduced IgE binding and a decreased capacity to induce mast cell degranulation, as demonstrated in vitro and in mouse models. Crucially, both variants can induce ​​Met e 1-specific IgG antibodies​​ (particularly IgG2a in mice) that act as ​​blocking antibodies​​, effectively inhibiting IgE from shrimp-allergic patients and sensitized mice from binding to the native allergen. This promotes immune tolerance and long-term desensitization, offering a promising preclinical therapeutic strategy for a common and potentially severe food allergy

Reference:

Wai et al. (2014). Immunization with Hypoallergens of Shrimp Allergen Tropomyosin Inhibits Shrimp Tropomyosin Specific IgE Reactivity. PLoS ONE 9(11): e111649. https://doi.org/10.1371/journal.pone.0111649

A hypoallergenic variant of Cyp c 1 was engineered through ​​site-directed mutagenesis​​ targeting the functional calcium-binding domains (CD and EF sites). Key aspartate residues at positions 51/53 (CD site) and 90/92 (EF site) were replaced with alanine to generate three mutants: Mut-CD, Mut-EF, and the double mutant Mut-CD/EF. The double mutant (Mut-CD/EF) exhibited ​​severe structural disruption​​, transitioning to a random coil conformation with minimal α-helical content, as confirmed by circular dichroism analysis.

​​Usage and Biology:​​

The hypoallergenic derivative Mut-CD/EF was designed for ​​allergen-specific immunotherapy (AIT)​​ of IgE-mediated fish allergy. Despite its disrupted conformation, it retained ​​immunogenicity​​: immunization of mice with Mut-CD/EF induced ​​IgG antibodies​​ that cross-reacted with wild-type parvalbumin and parvalbumins from various fish species. It represented a promising candidate for ​​safer AIT​​ due to its drastically reduced anaphylactic risk while maintaining potential for inducing blocking antibodies and immune tolerance.

​​Reference:​​

Swoboda et al. (2007). A Recombinant Hypoallergenic Parvalbumin Mutant for Immunotherapy of IgE-Mediated Fish Allergy. Journal of Immunology, 178(10), 6290–6296. https://doi.org/10.4049/jimmunol.178.10.6290

Design and Engineering Principle

The basic parts were inserted into the pET-32a vector, which features a T7 promoter for high-level expression, a His-tag for simplified purification, and a Trx tag to enhance soluble expression of the target allergen protein. The gene fragments encoding the engineered ​​hypoallergenic proteins​​ were first amplified via ​​PCR​​. The pET-32a vector was then digested with the corresponding ​​restriction enzymes​​ (BamHI and XhoI). Following purification, the target gene fragments and vector were assembled using ​​homologous recombination​​ (Figure 1).

Figure 1 Schematic diagram of the plasmid construction.

Cultivation, Protein Expression and Validation

​ As shown in Figure 2, PCR products and linearized vector were validated via agarose gel electrophoresis. Target gene fragments (330-720 bp) were confirmed for purity and accuracy through staining and UV visualization. Following gel extraction and purification, six allergen genes (mPhl p1, mBet v1, mDer p1, mDer p2, mCyp c1, mMet e1) were inserted into the linearized pET32a vector using homologous recombination technology, completing recombinant plasmid construction.

Figure 2 Agarose gel electrophoresis results of (A) gene amplification and (B) vector linearization.

The recombinant products were then transformed into E. coli Top 10 competent cells, and positive clones were selected on LB agar plates, respectively. Colony PCR and DNA sequencing verified the correct insertion and integrity of the allergen genes. Selected positive clones were sequenced, and the sequence data confirmed that each plasmid construct contained the correct allergen gene sequence with high fidelity (Figure 3). The validated plasmids were subsequently transformed into E. coli Origami (DE3) competent cells to enhance soluble expression of the recombinant hypoallergenic proteins.

Figure 3 (A) Transformation, (B) colony PCR and (C) Sequencing results of plasmids.

Next, we expanded the cultivation and used IPTG to induce the expression of the hypoallergenic protein derivatives. Our goal was to express and purify the recombinant allergen proteins for subsequent research. After cell lysis and centrifugation, we performed SDS-PAGE to identify the protein expression and purification conditions. The SDS-PAGE analysis results showed that all six hypoallergenic protein derivatives were successfully expressed, and the molecular weights were consistent with the expectations (Figure 4 A). Among them, mPhl p1, mBet v1, and mDer p1 were mainly expressed in the precipitate part, with less background proteins and high purity. These insoluble proteins were then denatured and refolded to obtain soluble proteins. For the three soluble-expressed proteins: mDer p2, mMet e1, and mCyp c1, we used nickel columns to purify them separately. The SDS-PAGE results indicated that we successfully obtained highly pure proteins (Figure 4 B).

Figure 4 SDS-PAGE results of (A) protein expression and (B) purification

Characterization

1. Protein quantitative analysis

Protein concentrations of both purified native allergens and their hypoallergenic derivatives were quantified using the Bradford assay to ensure accurate loading for subsequent IgE-binding affinity assays. A standard curve was constructed using bovine serum albumin (BSA) as a reference, exhibiting excellent linearity (R² = 0.9909) with the regression equation Y = 0.001403x + 0.004835 (Figure 5). All absorbance measurements at 595 nm were performed in triplicate, and sample concentrations were interpolated based on the standard curve (Table 2).

Figure 5 BSA standard curve test by Bradford assay.

(A) The absorbance values of the BSA standard samples. (B) Fitting results of the standard curve.

Table 2 Protein quantification results.

2. IgE affinity test of allergen protein

Next, we conducted IgE binding affinity analysis, the main purpose of which is to functionally evaluate the allergenic potential of natural and modified hypoallergenic derivatives, and fusion hypoallergenic derivatives. This assessment is crucial because the clinical severity of allergic reactions is largely determined by the affinity of the specific IgE antibodies against the allergen and its target epitope (Fromberg, 2006). High-affinity IgE antibodies can trigger strong activation of mast cells and basophils even in the presence of low concentrations of allergens, leading to significant release of inflammatory mediators and potential anaphylactic shock. In contrast, low-affinity IgE has significantly reduced ability to bind to the FcεRI receptor, thereby reducing the risk of severe allergic reactions (Udoye, Ehlers, & Manz, 2023). Therefore, by quantifying the degree of reduction in IgE binding affinity, it can provide a key indicator for in vitro experiments to determine whether our rational protein design strategy has successfully achieved the reduction of allergenicity.

The standard curve for human IgE quantification was generated using four-parameter logistic (4PL) regression in GraphPad Prism 9.5, demonstrating excellent fit (R² > 0.998), which validated the reliability of subsequent affinity measurements (Figure 6A). The IgE-binding capacities of six purified allergens—both native and hypoallergenic derivatives—were evaluated by ELISA. Coated proteins were incubated with 50 ng/mL human IgE. As shown in Figure 6B, significant reduction in IgE binding (p < 0.0001) was observed in four hypoallergenic derivatives compared to their native counterparts (mBet v1, mDer p2, mMet e1, mCyp c1). Although mPhl p1 and mDer p1 also showed decreased affinity, the reduction was more modest (p>0.05). These results confirmed the effectiveness of the rational design strategy in generating hypoallergenic proteins with markedly reduced IgE reactivity.

Figure 6 IgE affinity of allergen proteins. (A) IgE standard curve, (B) Comparison of natural allergens and hypoallergenic derivatives.

3. Cell proliferation experiment test

Building upon the IgE-binding affinity results, we further evaluated the cytotoxicity of both single and fusion hypoallergenic derivatives using a CCK-8 assay to assess their biosafety for potential in vivo therapeutic applications. This evaluation is essential to ensure that engineered biologics exhibit minimal adverse effects on cell viability before proceeding to animal studies or clinical use.

After incubating cells with each protein at concentrations of 1 nM, 10 nM, and 100 nM for 24 h and 48 h, significant differences in cytotoxicity profiles were observed. For mBet v1 and mCyp c1, at all tested concentrations and time points, there was almost no significant effect on cell viability, demonstrating extremely high biocompatibility (Figure 7B, F). For mPhl p1 and mMet e1, the toxicity showed a clear time and concentration dependence (Figure 7A, E). At low concentrations and short durations, the impact was minimal, but as the concentrations and durations increased, the toxicity significantly intensified. For mDer p1 and mDer p2, even at lower concentrations and shorter durations, it could cause a significant decrease in cell viability, indicating an inherent toxicity (Figure 7C, D). This initial verification has confirmed the biocompatibility of the hypoallergenic derivatives we used.

Figure 7 The results of the cell proliferation experiment.

Table 3 Part contribution about fusion derivatives

Part number	Part name	Part type	Contribution
BBa_25QXOB8J	F peptide (EFK8)	Basic part	New part
BBa_2596PY5I	pET32a-EFK8-PT-mPhl p1-GS-mBet v1-His tag	Composite part	New part
BBa_25K5E713	pET32a-EFK8-PT-mDer p1-GS-mDer p2-His tag	Composite part	New part
BBa_255LSZHN	pET32a-EFK8-PT-mMet e1-GS-mCyp c1-His tag	Composite part	New part
BBa_25H406D2	pET32a-EFK8-PT-EGFP-His tag	Composite part	New part

The peptide EFK8 (FEFEFKFK) is a self-assembling octapeptide engineered through de novo design, inspired by natural self-assembling motifs and amyloidogenic sequences observed in protein misfolding diseases.

Properties:

EFK8 is a ​​cationic amphipathic peptide​​ characterized by alternating phenylalanine (F) and glutamic acid (E)/lysine (K) residues. This sequence promotes spontaneous ​​self-assembly into β-sheet-rich nanofibers​​ under physiological conditions, driven by hydrophobic interactions (via phenylalanine stacks) and electrostatic forces (via Glu-Lys pairs).

Usage and Biology:

EFK8 is primarily utilized as a ​​self-assembling carrier or scaffold​​ in biomedical research, particularly in drug delivery, vaccine development, and tissue engineering. Its ability to form nanofibrous networks allows efficient encapsulation and presentation of antigens or therapeutics.

Reference:

Mohammed A, et al. 3D Networks from Self‐Assembling Ionic‐Complementary Octa‐Peptides. Macromolecular Symposia. Weinheim: WILEY‐VCH Verlag, 2007, 251(1): 88-95.

Design and Engineering Principle

To address the issue of multiple allergic reaction sensitivities and simplify the treatment plan, we have designed and manufactured a new type of fusion protein. This protein integrates multiple modified hypoallergenic derivatives that trigger similar immune responses into a single molecular structure. This strategy aims to achieve broad therapeutic effects, potentially avoiding the cumbersome process of multiple separate injections, thereby enhancing treatment efficiency and improving patient compliance. To achieve this, we investigated and selected the EFK8 self-assembling peptide (FEFEFKFK), which undergoes oligomerization driven by hydrophobic stacking of phenylalanine residues and electrostatic interactions of lysine residues. This process forms stable nanostructures that increase molecular size and reduce renal clearance, thereby prolonging the circulation half-life (Song et al., 2023).

We assembled short EFK8 oligonucleotide fragments firstly. Subsequently, we separately amplified single hypoallergenic derivative gene fragments with homologous arms through PCR. Then, we used overlapping extension PCR to splice these fragments together to generate the complete fusion allergen gene sequence. Finally, we performed restriction endonuclease digestion and ligation on the purified fusion gene and linearized vector backbone to obtain the desired recombinant plasmid product. Furthermore, we also constructed a plasmid that integrates the EFK8 peptide and the enhanced green fluorescent protein EGFP, which was used for the subsequent determination of the half-life (Figure 8).

Figure 8 Schematic diagram of the plasmid construction.

Cultivation, Protein Expression and Validation

As shown in Figure 9, PCR products were validated via agarose gel electrophoresis. Target gene fragments were confirmed for purity and accuracy through staining and UV visualization Figure 9A-B. Then, we used overlapping extension PCR to splice these fragments together to generate the complete fusion allergen gene sequence (Figure 9C). After the fragments were recovered through gel cutting, we used homologous recombination to recombine them.

Figure 9 Agarose gel electrophoresis results of gene amplification.

Next, we transferred the recombinant plasmid into the E. coli Top 10 bacteria, and then screen out the positive clones on the LB culture medium plate. The results of colony PCR and DNA sequencing indicated that we successfully obtained the plasmid with correct sequencing (Figure 10). These verified plasmids were subsequently transferred into E. coli Origami (DE3).

Figure 10 (A) Transformation, (B) colony PCR and (C) Sequencing results of plasmids.

We expanded the cultivation scale and used IPTG to induce the expression of the natural allergen proteins. The EFK8-mPhl p1-GS-mBet v1 (FPB) and EFK8-mDer p1-GS-mDer p2 (FDD) proteins are both expressed in the precipitate. Subsequently, a denaturation-reconstitution procedure is required to restore their soluble state. The EFK8-mMet e1-GS-mCyp c1 (FMC) protein was expressed in both the supernatant and the precipitate. We purified the soluble FMC protein using a nickel column. The SDS-PAGE results indicated that we were able to successfully purify this protein (Figure 11).

Figure 11 SDS-PAGE results of (A) protein expression and (B) purification.

Characterization

1. Protein quantitative analysis

Protein concentrations of purified fusion hypoallergenic derivatives were quantified using the Bradford assay to ensure accurate loading for subsequent IgE-binding affinity assays. A standard curve was constructed using bovine serum albumin (BSA) as a reference, exhibiting excellent linearity (R² = 0.9987) with the regression equation Y = 0.001563x + 0.002397 (Figure 12). All absorbance measurements at 595 nm were performed in triplicate, and sample concentrations were interpolated based on the standard curve (Table 4).

Figure 12 BSA standard curve test by Bradford assay of

(A) The absorbance values of the BSA standard samples. (B) Fitting results of the standard curve.

Table 4 Protein quantification results.

2. IgE affinity test of allergen protein

As illustrated in Figure 13A, the IgE standard curve demonstrated excellent linearity (R² = 0.9943), ensuring the reliability of the subsequent binding affinity quantification. Interestingly, comparative analysis revealed that two fusion allergens, ​​EFK8-mPhl p1-GS-mBet v1 (FPB)​​ and ​​EFK8-mDer p1-GS-mDer p2 (FDD)​​, exhibited significantly reduced IgE binding ratios compared to their respective single hypoallergenic constituents (mPhl p1 and mDer p1, respectively). ​​EFK8-mMet e1-GS-mCyp c1 (FMC)​​ also showed low IgE binding affinity, which was comparable to the levels observed for its individual hypoallergenic derivatives (mMet e1 and mCyp c1) (Figure 13B). This consistent reduction across all fusion constructs suggests that the fusion strategy effectively attenuates IgE immunoreactivity beyond the level achieved by individual engineered allergens, likely due to structural masking of key epitopes or altered antigen presentation.

Figure 13 IgE affinity of allergen proteins. (A) IgE standard curve, (B) Comparison of single and fusion hypoallergenic derivatives.

3. Cell proliferation experiment test

We evaluated the cytotoxicity profiles of both individual and fused hypoallergenic derivatives. For the fusion construct EFK8-PT-mPhl p1-GS-mBet v1, we observed that the intrinsic cytotoxicity of mPhl p1 influenced the overall profile of the fusion protein, leading to a detectable reduction in cell viability particularly under high-concentration or prolonged exposure conditions (Figure 14A-C).In contrast, the fusion protein EFK8-PT-mMet e1-GS-mCyp c1 exhibited a different behavior: although mMet e1 alone showed time- and concentration-dependent cytotoxicity, the fusion with mCyp c1—which itself displayed no appreciable toxicity—resulted in a composite molecule that did not induce significant cytotoxicity, even at high doses or after extended incubation (Figure 14D-F). Notably, the fusion protein EFK8-PT-mDer p1-GS-mDer p2 did not exhibit significant cytotoxicity under the tested conditions. This is in clear contrast to its individual components, both mDer p1 and mDer p2, which caused a marked decrease in cell viability, especially at higher concentrations (e.g., 100 nM) and with longer culture periods (48 h) (Figure 14G-I). These results suggest that the fusion strategy can alter the cytotoxic properties of the constituent hypoallergenic derivatives, in some cases mitigating the adverse effects observed with the individual components.

​ The CCK-8 results indicated that these fusion constructs (EFK8-PT-mDer p1-GS-mDer p2 and EFK8-PT-mMet e1-GS-mCyp c1) possessed excellent biological safety characteristics and had a negligible impact on cell viability. This highlighted that the fusion strategy not only can expand the coverage of immunotherapy but also enhance biocompatibility. These findings supported the advancement of the fusion of hypoallergenic derivatives towards further evaluation.

Figure 14 The results of the cell proliferation experiment.

4. Half-life test

To evaluate whether fusion with the self-assembling peptide EFK8 prolongs the half-life of hypoallergenic fusion proteins, we employed EGFP as a model protein incubated in fetal bovine serum at 37 °C for 5 days to simulate in vitro conditions. Fitting the fluorescence decay data to a one-phase exponential model revealed that EFK8-EGFP exhibited a significantly extended half-life (1.820 days) compared to EGFP alone (0.9819 days), nearly doubling its stability (Figure 15). This enhancement is likely attributable to EFK8-mediated oligomerization, which increases molecular size and reduces degradation rate.

Figure 15 Half-life of EGFP and EFK8-EGFP.

Fromberg, J. (2006). IgE as a marker in allergy and the role of IgE affinity. 61(10), 1234-1234. doi:https://doi.org/10.1111/j.1398-9995.2006.01222.x

Song, H., Su, Q., Nie, Y., Zhang, C., Huang, P., Shi, S., . . . Wang, W. (2023). Supramolecular assembly of a trivalent peptide hydrogel vaccine for cancer immunotherapy. Acta Biomaterialia, 158, 535-546. doi:https://doi.org/10.1016/j.actbio.2022.12.070

Udoye, C. C., Ehlers, M., & Manz, R. A. (2023). The B Cell Response and Formation of Allergenic and Anti-Allergenic Antibodies in Food Allergy. 12(12), 1501.

Wallner, M., Hauser, M., Himly, M., Zaborsky, N., Mutschlechner, S., Harrer, A., . . . Ferreira, F. (2011). Reshaping the Bet v 1 fold modulates TH polarization. Journal of Allergy and Clinical Immunology, 127(6), 1571-1578.e1579. doi:https://doi.org/10.1016/j.jaci.2011.01.064