Binding Proteins Hub Research | Innovations in Gut Microbiota and Binding Proteins

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May 28, 2025

28.05.2025

How to Fix a Gut Microbiome Ravaged by Antibiotics

A diet rich in diverse carbohydrates outperforms faecal transplants in mice at restoring microbial diversity, which has been linked to a range of health conditions

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May 27, 2025

27.05.2025

Recent Research Found a Surprising Link Between Coffee and Gut Health

Coffee is more than just a daily habit—it may play an active role in shaping a healthier gut. A recent large-scale, multi-cohort study has uncovered a strong and reproducible association between coffee consumption and the enrichment of Lawsonibacter asaccharolyticus, a gut microbe linked to anti-inflammatory effects. Using integrated multi-omic data and in vitro experiments, the researchers identified quinic acid—found in coffee—as a potential driver of this microbial response. This study offers compelling evidence of a direct biochemical connection between specific dietary components and beneficial shifts in the gut microbiome.

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May 26, 2025

26.05.2025

How Dietary Fiber May Help Disarm Cancer-causing Gut Bacteria

A groundbreaking new study uncovers how the food we eat can interact with the bacteria living in our gut - and potentially help trigger colon cancer. Scientists used a mouse model to explore the combined effects of diet, microbiome composition, and genetics on colorectal cancer risk. What they found is both sobering and hopeful.

Gut Dysbiosis Uncovered: How Gut Diversity & Gut Barrier Function Play a Crucial Role in Maintaining Your Health 

The human gut microbiota plays a pivotal role in maintaining overall health. When the composition and function of this microbial ecosystem become imbalanced, we talk about gut dysbiosis. This imbalance contributes to a dysregulated gut-immune axis, referring to impaired communication and feedback loop between the gut microbiota, intestinal barrier, and the immune system¹.

The Role of Binding Proteins in Gut Health

Gut health is essential for overall well-being, yet lifestyle habits, environmental factors such as diet, and medicines contribute to an increasing prevalence of gut dysbiosis and a compromised gut lining. These disruptions can have significant health implications, ranging from acute digestive discomfort to long-lasting challenges and broader systemic effects1. In fact, the U.S. microbiome has lost over 30% of its bacterial diversity — mainly due to antibiotics and poor diet2, 3.

Orally Delivered Toxin–Binding Protein Protects Against Diarrhoea in a Murine Cholera Model

The ongoing seventh cholera pandemic, which began in 1961, poses an escalating threat to public health. There is a need for new cholera control measures, particularly ones that can be produced at low cost, for the one billion people living in cholera-endemic regions. Orally delivered VHHs, functioning as target-binding proteins, have been proposed as a potential approach to control gastrointestinal pathogens. Here, we describe the development of an orally deliverable bivalent VHH construct that binds to the B-pentamer of cholera toxin, showing that it inhibits toxin activity in a murine challenge model. Infant mice given the bivalent VHH prior to V. cholerae infection exhibit a significant reduction in cholera toxin–associated intestinal fluid secretion and diarrhoea. In addition, the bivalent VHH reduces V. cholerae colonization levels in the small intestine by a factor of 10. This cholera toxin–binding protein holds promise for protecting against severe diarrhoea associated with cholera.

Allergenicity, Genotoxicity and Subchronic Toxicity Assessment of IgG Binding Protein LT Produced From Aspergillus oryzae

Gastrointestinal health is one of the fastest growing areas in the food and beverage industry, as its importance to overall healthand well-being is becoming increasingly recognized. Immunoglobulins play a key role in protecting the gastrointestinal tract,and nonbovine sources of immunoglobulins (including camel milk, which has a long history of consumption in East Africa andAsia) are increasing in popularity in Western countries as functional foods, particularly for individuals with allergies or intolerances to cow's milk. The physiological benefits of consuming certain heavy-chain immunoglobulins from camel milk relateto the binding domains of camelid single-domain antibodies; thus, a novel binding protein termed “immunoglobulin G (IgG)binding protein LT” (a dimer of two camelid single-domain antibody protein sequences) has been developed for use in food andbeverage products, to provide some of the physiological benefits attributed to consuming camel milk, on an industrial scale. Tosupport the safety of IgG binding protein LT for such use, a comprehensive safety assessment (in silico allergenicity assessment,in vitro genotoxicity studies [bacterial reverse mutation test and in vitro mammalian cell micronucleus test], and a 90-day gavagetoxicity study in rats) was conducted. The in silico allergenicity assessment results demonstrate that IgG binding protein LT ishighly unlikely to pose a risk of allergenic cross-reactivity, and there was no evidence of genotoxicity in vitro. There were no testarticle–related effects in the 90-day toxicity study. These data demonstrate the safety of IgG binding protein LT for its intendeduses in foods and beverages.

Fit-for-Purpose Heterodivalent Single-Domain Dntibody for Gastrointestinal Targeting of Toxin B from Clostridium Difficile

Single-domain antibodies (sdAbs), such as $\text{V}_\text{H}\text{Hs}$, are increasingly being developed for gastrointestinal (GI) applications against pathogens to strengthen gut health. However, what constitutes a suitable developability profile for applying these proteins in a gastrointestinal setting remains poorly explored. Here, we describe an in vitro methodology for the identification of sdAb derivatives, more specifically divalent $\text{V}_\text{H}\text{H}$ constructs, that display extraordinary developability properties for oral delivery and functionality in the GI environment. We showcase this by developing a heterodivalent $\text{V}_\text{H}\text{H}$ construct that cross-inhibits the toxic activity of the glycosyltransferase domains (GTDs) from three different toxinotypes of cytotoxin B (TcdB) from lineages of Clostridium difficile. We show that the $\text{V}_\text{H}\text{H}$ construct possesses high stability and binding activity under gastric conditions, in the presence of bile salts, and at high temperatures. We suggest that the incorporation of early developability assessment could significantly aid in the efficient discovery of $\text{V}_\text{H}\text{Hs}$ and related constructs fit for oral delivery and GI applications.

Protecting the Piglet Gut Microbiota Against ETEC-Mediated Post-Weaning Diarrhoea Using Specific Binding Proteins

Post-weaning diarrhoea (PWD) in piglets presents a widespread problem in industrial pig production and is often caused by enterotoxigenic E. coli (ETEC) strains. Current solutions, such as antibiotics and medicinal zinc oxide, are unsustainable and are increasingly being prohibited, resulting in a dire need for novel solutions. Thus, in this study, we propose and evaluate a protein-based feed additive, comprising two bivalent heavy chain variable domain ($V_{HH}$) constructs ($V_{HH}$-(GGGGS)$_3$-$V_{HH}$, BL1.2 and BL2.2) as an alternative solution to manage PWD. We demonstrate in vitro that these constructs bind to ETEC toxins and fimbriae, whilst they do no affect bacterial growth rate. Furthermore, in a pig study, we show that oral administration of these constructs after ETEC challenge reduced ETEC proliferation when compared to challenged control piglets (1-2 $\log_{10}$ units difference in gene copies and bacterial count/g faeces across day 2–7) and resulted in week 1 enrichment of three bacterial families (Prevotellaceae (estimate: 1.12 $\pm$ 0.25, $q$ = 0.0054), Lactobacillaceae (estimate: 2.86 $\pm$ 0.52, $q$ = 0.0012), and Ruminococcaceae (estimate: 0.66 $\pm$ 0.18, $q$ = 0.049)) within the gut microbiota that appeared later in challenged control piglets, thus pointing to an earlier transition towards a more mature gut microbiota. These data suggest that such $V_{HH}$ constructs may find utility in industrial pig production as a feed additive for tackling ETEC and reducing the risk of PWD in piglet populations.

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