Positive Effects of Adiponectin, BDNF, and GLP-1 on Cortical Neurons Counteracting Palmitic Acid Induced Neurotoxicity.

The prevalence of metabolic syndrome caused by diets containing excessive fatty acids is increasing worldwide. Patients with metabolic syndrome exhibit abnormal lipid profiles, chronic inflammation, increased levels of saturated fatty acids, impaired insulin sensitivity, excessive fat accumulation, and neuropathological issues such as memory deficits. In particular, palmitic acid (PA) in saturated fatty acids aggravates inflammation, insulin resistance, impaired glucose tolerance, and synaptic failure. Recently, adiponectin, brain-derived neurotrophic factor (BDNF), and glucose-like peptide-1 (GLP-1) have been investigated to find therapeutic solutions for metabolic syndrome, with findings suggesting that they are involved in insulin sensitivity, enhanced lipid profiles, increased neuronal survival, and improved synaptic plasticity. We investigated the effects of adiponectin, BDNF, and GLP-1 on neurite outgrowth, length, and complexity in PA-treated primary cortical neurons using Sholl analysis. Our findings demonstrate the therapeutic potential of adiponectin, BDNF, and GLP-1 in enhancing synaptic plasticity within brains affected by metabolic imbalance. We underscore the need for additional research into the mechanisms by which adiponectin, BDNF, and GLP-1 influence neural complexity in brains with metabolic imbalances.

Mental health effects associated with COVID-19 financial assistance in South Korea: a comparison of employment status.

BACKGROUND: COVID-19 has created tensions across different sectors of the society, but the impact has been unequal. Vulnerable people have been most affected, especially those with insecure employment and who have experienced economic hardships due to unemployment and lost wages. The combination of social change and economic hardships due to the pandemic increases the risk of poor mental health. Some countries have utilized financial assistance to alleviate economic hardships caused by COVID-19, and in South Korea, the central and local governments have implemented COVID-19 financial assistance. This study analysed the impact of financial assistance on mental health associated with working status during the COVID-19 pandemic in South Korea. METHODS: The participants of this study were randomly selected from residents of Gyeonggi-do after being proportionally allocated by resident registration population status. A total of 1,000 adult males and females aged 19 years or older in Gyeonggi-do who received financial assistance from the central and local governments were selected. A retrospective pre-post-study design was applied, and mental health surveys including the Patient Health Questionnaire-9 (PHQ-9), Generalized Anxiety Disorder 7-item scale (GAD-7) were applied. RESULTS: The results show that depression scores averaged 5.5 and anxiety scores averaged 4.4 before COVID-19 Financial Assistance. It is similar to the national average of 5.1 and 4.5 respectively at that time. After the assistance, depression scores dropped to 4.5, and anxiety scores dropped to 3.2. Before the assistance, depression and anxiety were higher among temporary day labourers with less job security, and they showed the most significant improvement in mental health. For full-time workers, there was no significant change in anxiety or depression after receiving the assistance. CONCLUSIONS: Financial assistance can provide material resources and also positively affect mental health. In particular, it had a greater impact on the relatively vulnerable groups, such as those in unstable employment.

Circular RNA circSMAD4 regulates cardiac fibrosis by targeting miR-671-5p and FGFR2 in cardiac fibroblasts.

Heart failure is a leading cause of death and is often accompanied by activation of quiescent cardiac myofibroblasts, which results in cardiac fibrosis. In this study, we aimed to identify novel circular RNAs that regulate cardiac fibrosis. We applied transverse aortic constriction (TAC) for 1, 4, and 8 weeks in mice. RNA sequencing datasets were obtained from cardiac fibroblasts isolated by use of a Langendorff apparatus and then further processed by use of selection criteria such as differential expression and conservation in species. CircSMAD4 was upregulated by TAC in mice or by transforming growth factor (TGF)-ß1 in primarily cultured human cardiac fibroblasts. Delivery of si-circSMAD4 attenuated myofibroblast activation and cardiac fibrosis in mice treated with isoproterenol (ISP). si-circSmad4 significantly reduced cardiac fibrosis and remodeling at 8 weeks. Mechanistically, circSMAD4 acted as a sponge against the microRNA miR-671-5p in a sequence-specific manner. miR-671-5p was downregulated during myofibroblast activation and its mimic form attenuated cardiac fibrosis. miR-671-5p mimic destabilized fibroblast growth factor receptor 2 (FGFR2) mRNA in a sequence-specific manner and interfered with the fibrotic action of FGFR2. The circSMAD4-miR-671-5p-FGFR2 pathway is involved in the differentiation of cardiac myofibroblasts and thereby the development of cardiac fibrosis.

Pan-cancer analysis reveals multifaceted roles of retrotransposon-fusion RNAs.

Transposon-derived transcripts are abundant in RNA sequences, yet their landscape and function, especially for fusion transcripts derived from unannotated or somatically acquired transposons, remains underexplored. Here, we developed a new bioinformatic tool to detect transposon-fusion transcripts in RNA-sequencing data and performed a pan-cancer analysis of 10,257 cancer samples across 34 cancer types as well as 3,088 normal tissue samples. We identified 52,277 cancer-specific fusions with ~30 events per cancer and hotspot loci within transposons vulnerable to fusion formation. Exonization of intronic transposons was the most prevalent genic fusions, while somatic L1 insertions constituted a small fraction of cancer-specific fusions. Source L1s and HERVs, but not Alus showed decreased DNA methylation in cancer upon fusion formation. Overall cancer-specific L1 fusions were enriched in tumor suppressors while Alu fusions were enriched in oncogenes, including recurrent Alu fusions in EZH2 predictive of patient survival. We also demonstrated that transposon-derived peptides triggered CD8+ T-cell activation to the extent comparable to EBV viruses. Our findings reveal distinct epigenetic and tumorigenic mechanisms underlying transposon fusions across different families and highlight transposons as novel therapeutic targets and the source of potent neoantigens.

Deimmunization of flagellin adjuvant for clinical application.

Flagellin is the cognate ligand for host pattern recognition receptors, toll-like receptor 5 (TLR5) in the cell surface, and NAIP5/NLRC4 inflammasome in the cytosol. TLR5-binding domain is located in D1 domain, where crucial amino acid sequences are conserved among diverse bacteria. The highly conserved C-terminal 35 amino acids of flagellin were proved to be responsible for the inflammasome activation by binding to NAIP5. D2/D3 domains, located in the central region and exposed to the outside surface of flagellar filament, are heterogeneous across bacterial species and highly immunogenic. Taking advantage of TLR5- and NLRC4-stimulating activities, flagellin has been actively developed as a vaccine adjuvant and immunotherapeutic. Because of its immunogenicity, there exist worries concerning diminished efficacy and possible reactogenicity after repeated administration. Deimmunization of flagellin derivatives while preserving the TLR5/NLRC4-mediated immunomodulatory activity should be the most reasonable option for clinical application. This review describes strategies and current achievements in flagellin deimmunization.

Colloidal inorganic nano- and microparticles for passive daytime radiative cooling.

Compared to traditional cooling systems, radiative cooling (RC) is a promising cooling strategy in terms of reducing energy consumption enormously and avoiding severe environmental issues. Radiative cooling materials (RCMs) reduce the temperature of objects without using an external energy supply by dissipating thermal energy via infrared (IR) radiation into the cold outer space through the atmospheric window. Therefore, RC has a great potential for various applications, such as energy-saving buildings, vehicles, water harvesting, solar cells, and personal thermal management. Herein, we review the recent progress in the applications of inorganic nanoparticles (NPs) and microparticles (MPs) as RCMs and provide insights for further development of RC technology. Particle-based RCMs have tremendous potential owing to the ease of engineering their optical and physical properties, as well as processibility for facile, inexpensive, and large area deposition. The optical and physical properties of inorganic NPs and MPs can be tuned easily by changing their size, shape, composition, and crystals structures. This feature allows particle-based RCMs to fulfill requirements pertaining to passive daytime radiative cooling (PDRC), which requires high reflectivity in the solar spectrum and high emissivity within the atmospheric window. By adjusting the structures and compositions of colloidal inorganic particles, they can be utilized to design a thermal radiator with a selective emission spectrum at wavelengths of 8-13 µm, which is preferable for PDRC. In addition, colloidal particles can exhibit high reflectivity in the solar spectrum through Mie-scattering, which can be further engineered by modifying the compositions and structures of colloidal particles. Recent advances in PDRC that utilize inorganic NPs and MPs are summarized and discussed together with various materials, structural designs, and optical properties. Subsequently, we discuss the integration of functional NPs to achieve functional RCMs. We describe various approaches to the design of colored RCMs including structural colors, plasmonics, and luminescent wavelength conversion. In addition, we further describe experimental approaches to realize self-adaptive RC by incorporating phase-change materials and to fabricate multifunctional RC devices by using a combination of functional NPs and MPs.

The anticancer effect of PASylated calreticulin-targeting L-ASNase in solid tumor bearing mice with immunogenic cell death-inducing chemotherapy.

L-Asparaginase (L-ASNase), a bacterial enzyme that degrades asparagine, has been commonly used in combination with several chemical drugs to treat malignant hematopoietic cancers such as acute lymphoblastic leukemia (ALL). In contrast, the enzyme was known to inhibit the growth of solid tumor cells in vitro, but not to be effective in vivo. We previously reported that two novel monobodies (CRT3 and CRT4) bound specifically with calreticulin (CRT) exposed on tumor cells and tissues during immunogenic cell death (ICD). Here, we engineered L-ASNases conjugated with monobodies at the N-termini and PAS200 tags at the C-termini (CRT3LP and CRT4LP). These proteins were expected to possess four monobody and PAS200 tag moieties, which did not disrupt the L-ASNase conformation. These proteins were expressed 3.8-fold more highly in E. coli than those without PASylation. The purified proteins were highly soluble, with much greater apparent molecular weights than expected ones. Their affinity (Kd) against CRT was about 2 nM, 4-fold higher than that of monobodies. Their enzyme activity (∼6.5 IU/nmol) was similar to that of L-ASNase (∼7.2 IU/nmol), and their thermal stability was significantly increased at 55 °C. Their half-life times were > 9 h in mouse sera, about 5-fold longer than that of L-ASNase (∼1.8 h). Moreover, CRT3LP and CRT4LP bound specifically with CRT exposed on tumor cells in vitro, and additively suppressed the tumor growth in CT-26 and MC-38 tumor-bearing mice treated with ICD-inducing drugs (doxorubicin and mitoxantrone) but not with a non-ICD-inducing drug (gemcitabine). All data indicated that PASylated CRT-targeted L-ASNases enhanced the anticancer efficacy of ICD-inducing chemotherapy. Taken together, L-ASNase would be a potential anticancer drug for treating solid tumors.

A conformation-selective protein binder for a KRAS mutant inhibits the interaction between RAS and RAF.

Small GTPases are key signaling nodes that regulate the cellular processes and subcellular events, and their abnormal activities and dysregulations are closely linked with diverse cancers. Here, we report the development of conformation-selective protein binders for a KRAS mutant. The conformation-specific protein binders were selected from a repebody scaffold composed of LRR (Leucine-rich repeat) modules through phage display and modular engineering against constitute active conformation of KRAS. Epitope of the selected binders was mapped to be located close to switch I of KRAS. The conformation-selective protein binders were shown to effectively block the interaction between active KRAS and RAS-binding domain of BRAF, suppressing the KRAS-mediated downstream signaling.

Unemployment risk of all employed working-age cancer survivors after cancer diagnosis in South Korea: a retrospective cohort analysis of population-based administrative data.

PURPOSE: As the cancer survival rate increases, discussions on the employment status of cancer survivors should be actively carried out worldwide. This study examined patients' working status changes after cancer diagnosis to provide the basis for cancer survivors' return to work. METHODS: We established a nationwide cohort to determine long-term work changes after cancer diagnosis. All patients aged 19 to 50 years who were newly diagnosed with cancer while working for the previous 3 years were considered as the case group in 2010. Using propensity score matching (PSM), the cases were matched by sex and age at a ratio of 1:3 with the control group. Kaplan-Meier curve and Cox hazard model analyses were performed to determine the occurrence of unemployment and return to work in the case and control groups on a yearly basis. RESULTS: According to the 6-year follow-up after cancer diagnosis, 26.5% of cancer patients and 23.2% of controls had lost their jobs by the end of the follow-up (P < .0001). Meanwhile, 50.5% of cancer patients and 57.4% of controls had returned to work after unemployment (P < .0001). Subsequently, based on the Cox hazard model, the unemployment risk of cancer patients was 1.42 times higher than that of the general population, while the proportion of those who returned to work was 1.15 times lower. CONCLUSION: Employment is very closely related to the quality of life of cancer survivors. These results highlight the need for a system that can support cancer survivors' work maintenance and return to work after unemployment during the treatment period and the fact that awareness of this must be improved.

Fever temperatures modulate intraprotein dynamics and enhance the binding affinity between monoclonal antibodies and the spike protein from SARS-CoV-2.

Fever is a typical symptom of most infectious diseases. While prolonged fever may be clinically undesirable, mild reversible fever (<39℃, 312 K) can potentiate the immune responses against pathogens. Here, using molecular dynamics and free energy calculations, we investigated the effect of febrile temperatures (38℃ to 40℃, 311 K to 313 K) on the immune complexes formed by the SARS-CoV-2 spike protein with two neutralizing monoclonal antibodies. In analyzing the conformational dynamics of the interactions between the antibodies and the spike protein under different thermal conditions, we found that, at mild fever temperatures (311-312 K), the binding affinities of the two antibodies improve when compared to the physiological body temperature (37℃, 310 K). Furthermore, only at 312 K, antibodies exert distinct mechanical effects on the receptor binding domains of the spike protein that may hinder SARS-CoV-2 infectivity. Enhanced antibody binding affinity may thus be obtained using appropriate temperature conditions.

Artificial intelligence for antibody reading comprehension: AntiBERTa.

Utilizing publicly available antibody big data resources, Leem et al. (2022) developed an antibody-specific language model, AntiBERTa, to understand the "language" of antibodies. Case studies reveal that AntiBERTa can be an extremely useful tool for antibody engineering.

Computational epitope binning reveals functional equivalence of sequence-divergent paratopes.

The therapeutic efficacy of a protein binder largely depends on two factors: its binding site and its binding affinity. Advances in in vitro library display screening and next-generation sequencing have enabled accelerated development of strong binders, yet identifying their binding sites still remains a major challenge. The differentiation, or "binning", of binders into different groups that recognize distinct binding sites on their target is a promising approach that facilitates high-throughput screening of binders that may show different biological activity. Here we study the extent to which the information contained in the amino acid sequences comprising a set of target-specific binders can be leveraged to bin them, inferring functional equivalence of their binding regions, or paratopes, based directly on comparison of the sequences, their modeled structures, or their modeled interactions. Using a leucine-rich repeat binding scaffold known as a "repebody" as the source of diversity in recognition against interleukin-6 (IL-6), we show that the "Epibin" approach introduced here effectively utilized structural modelling and docking to extract specificity information encoded in the repebody amino acid sequences and thereby successfully recapitulate IL-6 binding competition observed in immunoassays. Furthermore, our computational binning provided a basis for designing in vitro mutagenesis experiments to pinpoint specificity-determining residues. Finally, we demonstrate that the Epibin approach can extend to antibodies, retrospectively comparing its predictions to results from antigen-specific antibody competition studies. The study thus demonstrates the utility of modeling structure and binding from the amino acid sequences of different binders against the same target, and paves the way for larger-scale binning and analysis of entire repertoires.

Computational Identification and Design of Complementary ß-Strand Sequences.

The ß-sheet is a regular secondary structure element which consists of linear segments called ß-strands. They are involved in many important biological processes, and some are known to be related to serious diseases such as neurologic disorders and amyloidosis. The self-assembly of ß-sheet peptides also has practical applications in material sciences since they can be building blocks of repeated nanostructures. Therefore, computational algorithms for identification of ß-sheet formation can offer useful insight into the mechanism of disease-prone protein segments and the construction of biocompatible nanomaterials. Despite the recent advances in structure-based methods for the assessment of atomic interactions, identifying amyloidogenic peptides has proven to be extremely difficult since they are structurally very flexible. Thus, an alternative strategy is required to describe ß-sheet formation. It has been hypothesized and observed that there are certain amino acid propensities between ß-strand pairs. Based on this hypothesis, a database search algorithm, B-SIDER, is developed for the identification and design of ß-sheet forming sequences. Given a target sequence, the algorithm identifies exact or partial matches from the structure database and constructs a position-specific score matrix. The score matrix can be utilized to design novel sequences that can form a ß-sheet specifically with the target.

Deimmunization of flagellin for repeated administration as a vaccine adjuvant.

Flagellin, a protein-based Toll-like receptor agonist, is a versatile adjuvant applicable to wide spectrum of vaccines and immunotherapies. Given reiterated treatments of immunogenic biopharmaceuticals should lead to antibody responses precluding repeated administration, the development of flagellin not inducing specific antibodies would greatly expand the chances of clinical applications. Here we computationally identified immunogenic regions in Vibrio vulnificus flagellin B and deimmunized by simply removing a B cell epitope region. The recombinant deimmunized FlaB (dFlaB) maintains stable TLR5-stimulating activity. Multiple immunization of dFlaB does not induce FlaB-specific B cell responses in mice. Intranasally co-administered dFlaB with influenza vaccine enhanced strong Ag-specific immune responses in both systemic and mucosal compartments devoid of FlaB-specific Ab production. Notably, dFlaB showed better protective immune responses against lethal viral challenge compared with wild type FlaB. The deimmunizing B cell epitope deletion did not compromise stability and adjuvanticity, while suppressing unwanted antibody responses that may negatively affected vaccine antigen-directed immune responses in repeated vaccinations. We explain the underlying mechanism of deimmunization by employing molecular dynamics analysis.

Epitope-Paratope Interaction of a Neutralizing Human Anti-Hepatitis B Virus PreS1 Antibody That Recognizes the Receptor-Binding Motif.

Hepatitis B virus (HBV) is a global health burden that causes acute and chronic hepatitis. To develop an HBV-neutralizing antibody that effectively prevents HBV infection, we previously generated a human anti-preS1 monoclonal antibody (1A8) that binds to genotypes A-D and validated its HBV-neutralizing activity in vitro. In the present study, we aimed to determine the fine epitope and paratope of 1A8 to understand the mechanism of HBV neutralization. We performed alanine-scanning mutagenesis on the preS1 (aa 19-34, genotype C) and the heavy (HCDR) and light (LCDR) chain complementarity-determining regions. The 1A8 recognized the three residues (Leu22, Gly23, and Phe25) within the highly conserved receptor-binding motif (NPLGFFP) of the preS1, while four CDR residues of 1A8 were critical in antigen binding. Structural analysis of the epitope-paratope interaction by molecular modeling revealed that Leu100 in the HCDR3, Ala50 in the HCDR2, and Tyr96 in the LCDR3 closely interacted with Leu22, Gly23, and Phe25 of the preS1. Additionally, we found that 1A8 also binds to the receptor-binding motif (NPLGFLP) of infrequently occurring HBV. The results suggest that 1A8 may broadly and effectively block HBV entry and thus have potential as a promising candidate for the prevention and treatment of HBV infection.

Epitopes of Protein Binders Are Related to the Structural Flexibility of a Target Protein Surface.

Protein binders including antibodies are known not to bind to random sites of target proteins, and their functional effectiveness mainly depends on the binding region, called the epitope. For the development of protein binders with desired functions, it is thus critical to understand which surface region protein binders prefer (or do not prefer) to bind. The current methods for epitope prediction focus on static indicators such as structural geometry or amino acid propensity, whereas protein binding events are in fact a consequence of dynamic interactions. Here, we demonstrate that the preference for a binding site by protein binders is strongly related to the structural flexibility of a target protein surface. Molecular dynamics simulations on unbound forms of antigen structures revealed that the antigen surface in direct contact with antibodies is less flexible than the rest of the surface. This tendency was shown to be similar in other non-antibody protein binders such as affibody, DARPin, monobody, and repebody. We also found that the relatedness of epitopes to the structural flexibility of a target protein surface is dependent on the secondary structure elements of paratopes. Monobody and repebody, whose binding sites are composed of ß-strands, distinctively prefer to bind to a relatively more rigid region of a target protein. These observations enabled us to develop a simple epitope prediction method which shows a comparable performance to the commonly used ones.

Effects of Community-Based Interventions on Medication Adherence and Hospitalization for Elderly Patients with Type 2 Diabetes at Primary Care Clinics in South Korea.

Korean Disease Control and Prevention Agency launched Control and Prevention Community-based Registration and Management for Hypertension and Diabetes mellitus Project (CRMHDP) in Gwangmyeong city, 2009. This project has provided incentives on both patient and physician and has made private clinics and Public Health Center (PHC) in a community collaborate for effective chronic disease management among elderly people. This study aimed to evaluate the effects of CRMHDP on medication compliance and hospitalization due to diabetes-specific complications. The retrospective cohort study design was based on data of Korean National Health Insurance (KNHI) with 2 control areas (A & B) with usual primary care service similar to Gwangmyeong city regarding community health resources. The data on the study subjects were examined for the following 5 years since the baseline point. Medication adherence rates of CRMHDP-enrollees after the project was significantly higher than two control groups. For the hospitalization due to any complications, adjusted hazard ratio in the intervention group, compared to the control group A and B, were 0.76 (95% Confidence Interval: 0.65-0.78) and 0.52 (95% Confidence Interval 0.41-0.78), respectively. CRMHDP could successful in improving the management of type 2 diabetes mellitus among elderly people in South Korean primary care settings.

Computationally-guided design and affinity improvement of a protein binder targeting a specific site on HER2.

A protein binder with a desired epitope and binding affinity is critical to the development of therapeutic agents. Here we present computationally-guided design and affinity improvement of a protein binder recognizing a specific site on domain IV of human epidermal growth factor receptor 2 (HER2). As a model, a protein scaffold composed of Leucine-rich repeat (LRR) modules was used. We designed protein binders which appear to bind a target site on domain IV using a computational method. Top 10 designs were expressed and tested with binding assays, and a lead with a low micro-molar binding affinity was selected. Binding affinity of the selected lead was further increased by two-orders of magnitude through mutual feedback between computational and experimental methods. The utility and potential of our approach was demonstrated by determining the binding interface of the developed protein binder through its crystal structure in complex with the HER2 domain IV.

A Chimeric Antigen Receptor That Binds to a Conserved Site on MICA.

The NKG2D ligand MHC class I chain-related protein A (MICA) is expressed on many varieties of malignant cells but is absent from most normal tissues, and thus represents a potential target for chimeric Ag receptor (CAR) T cell-based therapeutics. However, there are more than 100 alleles of MICA, so the ability to target a conserved site is needed for a therapy to be used in most patients. In this study, we describe a fully human anti-MICA CAR created by fusing the single-chain fragment variable B2 to the full length DAP10 protein and the traditional CD3ζ signaling domain. Human T cells expressing the B2 CAR killed MICA-positive tumor cells, produced IFN-γ when in contact with MICA-positive tumor cells or plate-bound MICA protein, and inhibited PANC-1 growth in a mouse xenograft model. To localize B2's epitope on MICA, we used novel computational methods to model potential binding modes and to design mutational variants of MICA testing these hypotheses. Flow cytometry using a commercial anti-MICA/MICB Ab indicated that the variant proteins were expressed at high levels on transduced P815 cell lines. One variant protein (R38S/K40T/K57E) showed reduced staining with a B2-IgG1 fusion protein compared with controls and did not induce IFN-γ production by human T cells expressing the B2 CAR. These results show antitumor activity of MICA-specific CAR T cells and indicate an essential role for a conserved site in the exposed loop involving aa 38-57 of MICA. This study describes a novel MICA-specific CAR and discusses its potential use as a cancer therapeutic.

Generation and Characterization of Monoclonal Antibodies to the Ogawa Lipopolysaccharide of Vibrio cholerae O1 from Phage-Displayed Human Synthetic Fab Library.

Vibrio cholerae, cause of the life-threatening diarrheal disease cholera, can be divided into different serogroups based on the structure of its lipopolysaccharide (LPS), which consists of lipid-A, corepolysaccharide and O-antigen polysaccharide (O-PS). The O1 serogroup, the predominant cause of cholera, includes two major serotypes, Inaba and Ogawa. These serotypes are differentiated by the presence of a single 2-O-methyl group in the upstream terminal perosamine of the Ogawa O-PS, which is absent in the Inaba O-PS. To ensure the consistent quality and efficacy of the current cholera vaccines, accurate measurement and characterization of each of these two serotypes is highly important. In this study, we efficiently screened a phage-displayed human synthetic Fab library by bio-panning against Ogawa LPS and finally selected three unique mAbs (D9, E11, and F7) that specifically react with Ogawa LPS. The mAbs bound to Vibrio cholerae vaccine in a dose-dependent fashion. Sequence and structure analyses of antibody paratopes suggest that IgG D9 might have the same fine specificity as that of the murine mAbs, which were shown to bind to the upstream terminal perosamine of Ogawa O-PS, whereas IgGs F7 and E11 showed some different characteristics in the paratopes. To our knowledge, this study is the first to demonstrate the generation of Ogawa-specific mAbs using phage display technology. The mAbs will be useful for identification and quantification of Ogawa LPS in multivalent V. cholerae vaccines.