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We previously reported a chromatography system for purifying immunoglobulin M (IgM) using N,N,N',N'-ethylenediaminetetrakis(methylenephosphonic acid)-modified zirconia particles that selectively absorb immunoglobulins. Here, we report a simple procedure for preparing biotinylated IgM from hybridoma culture medium using this zirconia-based chromatography system. The culture medium of an IgM-producing hybridoma cell line was used as the starting sample solution, and the IgM in the medium was concentrated and partially purified by zirconia chromatography. Next, 9-(biotinamido)-4,7-dioxanonanoic acid N-succinimidyl ester was added to react with the proteins in the sample. Subsequently, only the biotinylated IgM was isolated by Capto Core 400 polishing column chromatography. The entire process was easy to perform, could be completed within 2 h, and provided highly pure biotin-labeled IgM. This procedure is expected to be applicable to the labeling of IgM with various compounds and drugs.
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We herein report a case of sporadic intra-abdominal desmoid-type fibromatosis in which contrast-enhanced ultrasonography (US) combined with superb microvascular imaging (SMI) was useful for preoperative diagnosis. 18-Fluorodeoxyglucose positron emission tomography performed for systematic screening for lung cancer revealed an abnormal accumulation in the abdominal cavity. Transabdominal US showed a tumor with a mixture of hypoechoic and hyperechoic areas. Contrast-enhanced US combined with SMI revealed dendritic blood flow signals and no abnormal vascular network within the tumor. Macroscopic examination of the resected specimen revealed a white tumor with relatively clear boundaries. Microscopic examination revealed spindle cells with poor atypia proliferating in bundles with collagenous stromal cells. Immunohistochemistry showed nuclear localization of beta-catenin within the tumor cells. From these findings, we finally diagnosed intra-abdominal desmoid-type fibromatosis. Contrast-enhanced US combined with SMI is useful for diagnosing intra-abdominal desmoid-type fibromatosis.
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A 23-year-old man with a history of anemia of unknown cause was referred to our hospital. He had experienced melena three times: at 6 months, 10 years, and 20 years of age. He underwent upper and lower gastrointestinal endoscopy at 10 and 20 years of age, and small intestinal capsule endoscopy twice at 20 years of age, but the site of bleeding could not be identified. At first, a transabdominal ultrasound was performed for suspected Meckel's diverticulum. A cystic diverticulum was found in the ileum, with an apertural diameter of 5 mm and a total size of 4 cm. The cyst showed an area of loss of wall stratification, which appeared to be an ulcer scar. Based on these observations, we diagnosed anemia resulting from a hemorrhagic Meckel's diverticulum and performed laparoscopic resection. Postoperative histopathology revealed ectopic gastric mucosa and ulcer formation within the Meckel's diverticulum, which was thought to be the cause of the bleeding. Meckel's diverticulum should be considered in cases of hemorrhage in young patients. A transabdominal ultrasound as a screening test detected a diverticulum with an ulcer scar in the ileum, which led to the identification of the underlying disease.
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PURPOSE: Osteosclerotic images are known as an image appearance of occult femoral neck fractures in X-ray images. The aim of this study is to investigate frequency enhancement processing that improves the visibility of the osteosclerotic image. METHODS: We acquired three sclerotic bone images with different thicknesses, and self-made bone equivalent phantoms were set up on a pelvic phantom. The frequency processing type and high-density enhancement coefficients were applied to the X-ray images taken at RF-A(1.0, 2.0), C(2.0, 4.0), D(1.0), and H(2.0, 4.0). For the physical index, we compared the difference in signal values between the sclerotic and background normal bone. We evaluated the preference using Scheffé's paired comparison methods for the visual index. RESULTS: For the physical index, RF-C(4.0) had the most significant signal value difference for all 3 bone stiffness images. For the visual index, RF-C(4.0) showed the highest preference. CONCLUSION: Using frequency-enhanced processing, RF-C(4.0) was suggested to improve the visibility of the osteosclerosis image.
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Fracturas del Cuello Femoral , Fantasmas de Imagen , Fracturas del Cuello Femoral/diagnóstico por imagen , Humanos , Osteosclerosis/diagnóstico por imagen , Fracturas Cerradas/diagnóstico por imagen , Intensificación de Imagen Radiográfica/métodosRESUMEN
A 74-year-old man presented to the emergency department with the chief complaint of abdominal pain. A computed tomography scan showed paralytic ileus. An ileostomy tube was placed, but the symptoms of bowel obstruction did not improve. Two days after admission, the patient's renal function deteriorated. Transabdominal ultrasound (TUS) showed linear high-intensity echoes consistent with a fibrotic band and microbubbles suggestive of circulatory disturbance in the dilated intestinal tract. Subsequent contrast-enhanced ultrasound revealed circulatory disturbance of the small bowel wall. Emergency surgery was performed under the diagnosis of strangulated ileus. Intraoperative examination revealed that the terminal ileum was strangulated by a fibrotic band from the retroperitoneum, which was confirmed by TUS. The fibrotic band was resected, the strangulation was released, and ileocecal resection was performed. Postoperatively, intestinal peristalsis was rapidly restored. TUS was able to depict the fibrotic band, which could not be detected by a computed tomography scan, allowing the patient to undergo immediate surgical treatment. We herein report this case of strangulated bowel obstruction in which TUS and contrast-enhanced ultrasound were useful in preoperative assessment of the patient's condition.
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We report a patient with a mucocele with diffuse wall thickening diagnosed by transabdominal ultrasonography and contrast-enhanced ultrasonography. Transabdominal ultrasonography showed diffuse thickening of the entire appendix wall and an anechoic area that appeared to be fluid collected throughout the appendix lumen. However, the "onion skin sign" was not detected. Contrast-enhanced ultrasonography combined with superb microvascular imaging revealed abundant mucosal blood flow and no abnormal vascular network within the mucosa of the appendix wall. We preoperatively diagnosed a mucocele complicated by acute and chronic appendicitis, and ileocecal resection was performed. Macroscopic and microscopic findings of the resected specimens demonstrated that the appendiceal wall was diffusely thickened, with fibrosis and inflammatory cell infiltration, and that the appendiceal root rumen was narrowed with epithelial hyperplasia. No neoplastic changes were observed. The cause of the appendiceal mucocele was likely fibrosis and stenosis at the root of the appendix due to initial acute appendicitis.
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Due to the scarcity of large-sized prospective databases, the Japanese Joint Committee for Lung Cancer Registry conducted a nationwide prospective registry for newly diagnosed and untreated pleural mesothelioma. All new cases diagnosed pathologically as any subtype of pleural mesothelioma in Japan during the period between April 1, 2017, to March 31, 2019, were included before treatment. Data on survival were collected in April 2021. The eligible 346 patients (285 men [82.3%]; 61 women [17.7%]; median age, 71.0 years [range, 44-88]) were included for analysis. Among these patients, 138 (39.9%) underwent surgery, 164 (47.4%) underwent non-surgical therapy, and the remaining 44 (12.7%) underwent best supportive care. The median overall survival for all 346 patients was 19.0 months. Survival rates at 1, 2, and 3 years for all patients were, 62.8%, 42.3%, and 26.5%, respectively. Median overall survival was significantly different among patients undergoing surgery, non-surgical treatment, and best supportive care (32.2 months vs. 14.0 months vs. 3.8 months, p < 0.001). The median overall survival of patients undergoing pleurectomy/decortication and extrapleural pneumonectomy was 41.8 months and 25.0 months, respectively. Macroscopic complete resection resulted in longer overall survival than R2 resection and partial pleurectomy/exploratory thoracotomy (41.8 months vs. 32.2 months vs. 16.8 months, p < 0.001). Tumor shape, maximum tumor thickness, and sum of three level thickness were significant prognostic factors. The data in the prospective database would serve as a valuable reference for clinical practice and further studies for pleural mesothelioma.
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Neoplasias Pulmonares , Mesotelioma Maligno , Mesotelioma , Neoplasias Pleurales , Masculino , Humanos , Femenino , Anciano , Japón/epidemiología , Resultado del Tratamiento , Mesotelioma/epidemiología , Mesotelioma/terapia , Neoplasias Pleurales/epidemiología , Neoplasias Pleurales/terapia , Neoplasias Pulmonares/epidemiología , Neoplasias Pulmonares/terapia , Estudios RetrospectivosRESUMEN
We previously generated three types of anti-glycan monoclonal IgM antibodies that react with certain structures on the glycans of glycosphingolipids and glycoproteins. As the nucleotide sequences for the variable regions of these IgM antibodies showed homology with those of anti-DNA antibodies deposited in public databases, we analyzed the reactivity of the anti-glycan IgM antibodies to DNA by ELISA. We found that anti-α2,6-sialyl LacNAc IgM in the supernatant of a hybridoma culture cross-reacted with DNA, and after purification of the IgM by zirconia column chromatography, the highly purified IgM showed increased cross-reactivity to DNA. As most of the contaminating bovine serum proteins in the culture supernatant were removed by the purification process, it is likely that a part of the removed components influences antibody reactivity to DNA. Purified anti-DNA antibodies prepared from lupus model NZB/W F1 and MRL/lpr mouse sera and normal human serum were then analyzed, and similar results showing increased reactivity to DNA were obtained. Furthermore, ELISA using these purified antibodies and various carbohydrate antigens showed that the antigen-binding specificity of these antibodies was altered by the purification process from serum-containing antibody preparations. Our results indicate that mammalian serum contains components that strongly influence antibody reactivity to carbohydrate antigens, including DNA.
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Anticuerpos Antinucleares , Polisacáridos , Ratones , Humanos , Animales , Ratones Endogámicos MRL lpr , Ensayo de Inmunoadsorción Enzimática , Inmunoglobulina M , ADN , MamíferosRESUMEN
Immobilized metal ion affinity chromatography (IMAC) is one of the most common purification techniques for histidine-tagged proteins (His-tagged proteins). IMAC enables the purification of His-tagged proteins at high purity on the basis of coordination bonds between His-tags and metal ions (such as Ni2+, Co2+, and Cu2+) immobilized on the matrices in columns. However, IMAC requires low-pH solutions or high-concentration imidazole solutions for eluting His-tagged proteins, which can affect protein conformation and activity. The present study provides a His-tagged protein purification method using zirconia particles modified with phosphate groups. This method is based on the electrostatic attractions between a His-tag moiety of proteins and phosphate groups on the zirconia particles; this method requires only high-concentration salt solutions at pH 7.0 for eluting the proteins. A column packed with phosphate-modified zirconia particles was demonstrated to enable the purification of two model His-tagged proteins-His-tagged green fluorescent protein and His-tagged alkaline phosphatase fused with maltose binding protein. Thus, this chromatography method is useful for purifying His-tagged proteins without any pH stress or additives. Additionally, because of the mechanical properties of the zirconia particles, this technique enables high-performance purification at a high flow rate.
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Histidina , Fosfatos , Histidina/química , Indicadores y Reactivos , Cromatografía de Afinidad/métodos , Proteínas RecombinantesRESUMEN
Eight workers involved in packing cross-linked water-soluble acrylic acid polymer, an organic substance, developed pulmonary fibrosis, and the upper lobe was the most affected. The dust concentration in the polymer packing workstation was measured. Chest computed tomography (CT) was obtained for 82 individuals, including the 8 workers mentioned above. Three workers were histopathologically examined. In six of these eight workers, central pulmonary fibrosis and secondary bulla formation caused pneumothorax. Histopathologically, multiple centrilobular fibrotic foci were observed. Chest CT revealed centrilobular nodular opacity and interlobular septal thickening, suggesting early lesions in the workers because the dust concentration was remarkably high. Although the pathogenesis of the disease is unclear, we reported the occurrence of pulmonary fibrosis caused by the exposure to cross-linked water-soluble acrylic acid polymers in humans as it has not been reported earlier.
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Fibrosis Pulmonar , Trastornos Respiratorios , Enfermedades Respiratorias , Humanos , Fibrosis Pulmonar/patología , Polímeros , Pulmón/patología , Enfermedades Respiratorias/patología , Trastornos Respiratorios/patología , PolvoRESUMEN
High-performance and cost-effective purification is necessary for the development of antibody drugs. This study found that nanoparticles of zirconia modified with phosphate groups selectively adsorb immunoglobulin G (IgG) antibodies against serum proteins with high adsorption capacity. The IgG antibodies collected from the zirconia nanoparticle surfaces retain their molecular conformation. Importantly, zirconia nanoparticles have the highest affinity for human IgG antibodies among tested mammalian IgG antibodies. The affinity for human IgG subclasses is in the order IgG3 > IgG1 > IgG2, which contrasts with a conventional ligand (Protein A) that has a lower affinity for IgG3. Because zirconia nanoparticles are chemically and mechanically stable, they can be utilized for the purification of antibody drugs not only in batch methods but also in chromatography as a process upstream or downstream of Protein A chromatography and even as an alternative process.
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Inmunoglobulina G , Fosfatos , Animales , Humanos , Adsorción , MamíferosRESUMEN
Immunization of mice with liposomes consisting of dipalmitoylphosphatidylcholine, cholesterol, lipid A, and glycosphingolipids (GSLs) can efficiently induce the production of antibodies that recognize specific GSLs. Here, we analyzed the effect of GSL species on the particle sizes of GSL-containing liposomes. We prepared liposomes containing Gb4Cer/globoside, GM3, and several artificial GSLs, and analyzed their particle sizes in phosphate-buffered saline by dynamic light scattering. The particle sizes of liposomes were significantly altered by the addition of GSLs, and they formed 65- to 1737-nm particle sizes depending on their constituent GSL species. We compared the sizes of each GSL-containing liposome with the IgM- or IgG-inducing activity of these liposomes in mice, and found a positive correlation between increasing liposome size and IgG-inducing activity. We also determined the nucleotide sequences of the heavy and light chain variable regions of anti-Gb4Cer IgM and IgG3 obtained from the Gb4Cer-containing liposome-immunized mice, and found that they were composed of different gene segments. This result indicates that the GSL-containing liposomes induce the production of IgG3 through an immune pathway different from that of IgM, rather than efficiently inducing class switching.
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Glicoesfingolípidos , Liposomas , Ratones , Animales , Inmunoglobulina M , Inmunoglobulina GRESUMEN
We herein report a case of an idiopathic intramural hematoma in the gastric wall that presented with symptoms of anemia. Esophagogastroduodenoscopy revealed a submucosal tumor-like lesion on the anterior gastric wall below the gastric fundus. Noncontrast-enhanced ultrasonography showed an anechoic area with indistinct boundaries mainly located within the submucosal layer in the gastric wall, and the lesion showed no contrast enhancement on contrast-enhanced ultrasonography. Based on a comprehensive analysis of the above-mentioned ultrasonographic imaging findings, a final diagnosis of idiopathic intramural gastric hematoma was made. The patient was treated conservatively, and changes in the size and internal ultrasonographic characteristics of the mass were followed up by ultrasonography. Six months later, esophagogastroduodenoscopy confirmed that the mass had disappeared.
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By using EDTPA-modified zirconia particles that selectively adsorb immunoglobulins in a column, we developed a chromatography separation system for efficient concentrating and purifying of IgM from hybridoma culture supernatants. Hybridoma culture supernatants containing IgMs were diluted 3-fold with 10 mM phosphate buffer (pH 7.0) and passed through the column. During this process, zirconia particles selectively adsorbed these IgMs, and most of the contaminating proteins flowed out into the flow-through. The adsorbed IgMs were easily eluted with a small volume of 400 mM phosphate buffer (pH 8.0), and high-concentration IgM solutions were prepared. Subsequent simple processing using a Capto™ Core 400 cartridge column provided highly purified IgM. The operation is easy, and the activity of IgM is maintained because the purification process is performed using only neutral ranges of phosphate buffers. Here, we showed that anti-globoside and anti-CDw75 IgM purified by this method can be used to stain cervical cancer and Burkitt lymphoma cells that specifically express these respective tumor-associated carbohydrate antigens.
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Anticuerpos Monoclonales , Fosfatos , Cromatografía de Afinidad/métodos , Electroforesis en Gel de Poliacrilamida , Hibridomas , Inmunoglobulina M/química , CirconioRESUMEN
BACKGROUND: Chest radiography (CR) is employed as the evaluation of pneumoconiosis; however, we sometimes encounter cases in which computed tomography (CT) is more effective in detecting subtle pathological changes or cases in which CR yields false-positive results. PURPOSE: To compare CR to CT in the diagnosis of early-stage pneumoconiosis. MATERIAL AND METHODS: CR and CT were performed for 132 workers with an occupational history of mining. We excluded 23 cases of arc-welder's lung. Five readers who were experienced chest radiologists or pulmonologists independently graded the pulmonary small opacities on CR of the remaining 109 cases. We then excluded 37 cases in which the CT data were not sufficient for grading. CT images of the remaining 72 cases were graded by the five readers. We also assessed the degree of pulmonary emphysema in those cases. RESULTS: The grade of profusion on CR (CR score) of all five readers was identical in only 5 of 109 cases (4.6%). The CR score coincided with that on CT in 40 of 72 cases (56%). The CT score was higher than that on CR in 13 cases (18%). On the other hand, the CT score was lower than that on CR in 19 cases (26%). The incidence of pulmonary emphysema was significantly higher in patients whose CR score was higher than their CT score. CONCLUSION: CT is more sensitive than CR in the evaluation of early-stage pneumoconiosis. In cases with emphysema, the CR score tends to be higher in comparison to that on CT.
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Neumoconiosis , Enfisema Pulmonar , Polvo , Humanos , Pulmón/diagnóstico por imagen , Pulmón/patología , Neumoconiosis/diagnóstico por imagen , Neumoconiosis/patología , Enfisema Pulmonar/diagnóstico por imagen , Radiografía Torácica , Tomografía Computarizada por Rayos X/métodosRESUMEN
The requirement for compensation for diffuse pleural thickening in benign asbestos pleural effusion include five computed tomography findings of organized pleural effusion: [1] heterogeneity in the pleural effusion, [2] declined chest capacity, [3] "crow's feet" sign at the pleura, [4] immobilization of effusion volume, and [5] air in the effusion. Pleural effusion is diagnosed as organized, immobilized, and in the state of diffuse pleural thickening if at least three of these items are fulfilled, ([1] and [3] compulsory + one of the remaining items). This retrospective study investigated whether the requirement to confirm no organized pleural effusion changes after a follow-up of >3 months were available for cases fulfilling three of the five items; i.e., the confirmation of only [2] with [1] and [3]. Of 302 cases recognized by the Japanese laws, 105 cases with diffuse pleural thickening with organized effusion were enrolled. The number of subjects who fulfilled the diagnostic requirement for organized pleural effusion was confirmed. Eight subjects had a full score of 5 points, 82 subjects scored 4 points, and only 15 subjects scored 3 points. Furthermore, no changes were observed in the organized pleural effusion volume after a follow-up of >3 months.
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Amianto , Asbestosis , Enfermedades Pleurales , Derrame Pleural , Amianto/efectos adversos , Humanos , Japón , Enfermedades Pleurales/diagnóstico por imagen , Derrame Pleural/diagnóstico por imagen , Estudios RetrospectivosRESUMEN
We report 3 Japanese cases with increased colonic 18-fluorodeoxyglucose (FDG) uptake in association with melanosis coli. Case 1: A 56-year-old woman received 18-FDG positron emission tomography (PET) for comprehensive medical checkup. Case 2: A 79-year-old man received FDG-PET for follow-up evaluation for intestinal cancer. Case 3: A 51-year-old woman received FDG-PET for medical checkup. Each showed increased cecal and/or ascending colonic uptake without any colonic wall changes. Colonoscopy detected melanosis coli where FDG uptake was demonstrably increased. Neither malignancy nor inflammatory response was confirmed. Succeeding follow-ups showed neither malignant nor inflammatory lesions in any of the patients.
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Malignant myoepithelioma of the scrotum is extremely rare. We report the case of a 51-year-old man with malignant myoepithelioma of the scrotum, wherein computed tomography and magnetic resonance imaging revealed a lobulated soft tissue mass with calcification, cystic component, and solid component with gradual contrast enhancement on dynamic contrast-enhanced scans. The patient presented with scrotal induration, and there was no elevation of tumor markers and no evidence of a metastatic lesion on computed tomography and magnetic resonance imaging. Histopathological examination of the resected scrotal specimen confirmed a well-circumscribed solid tumor with septa, a small area of hemorrhage, and necrosis. The subsequent diagnosis was malignant myoepithelioma of the scrotum. This case shows that scrotal malignant myoepithelioma might appear as a well-defined lobulated mass with cystic regions. We conjecture that the enhancement pattern and apparent diffusion coefficient values can be potential markers for scrotal myoepithelial tumors.
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Eosinophilic gastroenteritis (EGE) can present findings on computed tomography (CT) images that resemble malignant tumors. EGE is generally treated with systemic oral steroid administration, which is reportedly effective in relieving symptoms at least temporarily. Here, we report a case of EGE that mimicked malignant lymphoma in a gastroduodenal lesion, in which treatment with a potassium-competitive acid blocker without systemic oral steroid administration relieved the symptoms and reversed the initial image findings. A 56-year-old woman became aware of discomfort in her epigastric region, which gradually worsened. This case showed antroduodenal wall thickness, which mimicked a malignant lymphoma with increased F-18 fluorodeoxyglucose (FDG) uptake by positron emission tomography/computed tomography (PET/CT). An upper gastrointestinal endoscopy revealed protruding erosions in the antrum and severe edematous changes in the duodenum. Extensibility of the normal gastric wall by insufflation also occurred. However, an abdominal ultrasound indicated wall thickening with a preserved wall structure. Histological examinations showed a large amount of eosinophil infiltration, mainly in the gastroduodenal mucosa, without malignant cells. Thus, we made a final diagnosis of EGE. The patient was then administered 20 mg vonoprazan fumarate per day because she refused steroid treatment and because the pathological lesion was also localized from the duodenum to the antrum. After 3 weeks of potassium-competitive acid blocker treatment, her symptoms were alleviated, and the degree and extent of thickening of the gastroduodenal wall in upper gastrointestinal endoscopy and ultrasonographic image findings had remarkably improved.
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We previously reported an antibody (clone ID: FR9, IgM-κ) that recognizes the sialyl oligosaccharide Neu5Acα2,6Galß1,4GlcNAc as an epitope on glycoproteins and glycolipids. In the present study, we developed an antibody (clone ID: AFR45, IgM-κ) that recognizes Neu5Acα2,3Galß1,4GlcNAc/Glc as an epitope on glycoproteins and glycolipids and compared the nucleotide and amino acid sequences of the immunoglobulin gene variable regions with those of FR9. The heavy chain variable (VH) regions of FR9 and AFR45 were encoded by different VH gene segments, each of which was composed of a characteristic D gene segment. The major differences between VH genes encoding various antibodies deposited in public databases and FR9 and AFR45 were identified in the D gene segment, indicating that D genes play a critical role in determining the epitope specificity of these antibodies. Surprisingly, although FR9 and AFR45 were obtained independently from different mice immunized with different immunogens, the light chain variable (VL) region nucleotide sequences were identical. The VL gene consisted of Igkv4-57 and Igkj4 gene segments (Igkv4-57j4), the sequences of which were identical to VL genes for a number of antibodies against meningococcal group C capsular polysaccharide deposited in public databases. As this polysaccharide is a sialic acid homopolymer, these results indicate that Igkv4-57j4 encodes a VL common to immunoglobulins that recognize sialylated glycans.