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Reconstituted Membrane Systems for Assaying Membrane Proteins in Controlled Lipid Environments
Published in Qiu-Xing Jiang, New Techniques for Studying Biomembranes, 2020
Voltage-sensitive dyes can measure the change in transmembrane electrostatic potential. They can be called potential-sensitive probes. These dyes change their absorption or emission fluorescence as a function of transmembrane potential. This method was started by Lawrence Cohen in the middle of 1970s when he used squid giant axons to screen available dyes,62–64 and later developed several of them, such as diASP, di-4-ANEPPS,65–67 di-8-ANEPPS,68 and di-4-ANEPPDHQ.69 Due to their chromophores, these dyes are classified as hemocyanin. An array of different potential-sensitive dyes were later developed in order to investigate varying aspects of neuronal functions in real time.
For The Want of a Nail … Trace Elements in Health and Disease
Published in Owen M. Rennert, Wai-Yee Chan, Metabolism of Trace Metals in Man, 2017
Paul Saltman, Jack Hegenauer, Linda Strause
Oxygen transport and exchange is carried out by invertebrates using the copper protein, hemocyanin. This bright blue respiratory pigment provides O2 exchange from circulation to tissues. No trace elements other than iron and copper have been related to oxygen transport. Zinc, however, is an integral part of the enzyme carbonic anhydrase, which facilitates gas exchange in blood and tissues by forming dissolved carbonic acid from gaseous carbon dioxide and water.
Recent Advances in Diagnosis and Management of Shellfish Allergy
Published in Andreas L. Lopata, Food Allergy, 2017
Sandip D. Kamath, Roni Nugraha, Andreas L. Lopata
Most of the clinical studies on cross-reactivity have been conducted using tropomyosin as the major pan-allergen. However, other shellfish allergens may play a role in immunological cross- sensitisation. A recent study has shown that allergens other than tropomyosin, such as arginine kinase might also be responsible for seafood-mite cross-reactivity (Marinho et al. 2006, Gamez et al. 2014). Hemocyanin has been demonstrated to play a role in seafood-mite sensitivity as well as being characterised as a cockroach allergen (Giuffrida et al. 2014, Khurana et al. 2014).
Porphyromonas gingivalis diffusible signaling molecules enhance Fusobacterium nucleatum biofilm formation via gene expression modulation
Published in Journal of Oral Microbiology, 2023
Yukiko Yamaguchi-Kuroda, Yuichiro Kikuchi, Eitoyo Kokubu, Kazuyuki Ishihara
Based on the obtained gene sequence, an antibody against the protein was prepared. The peptide ESEVKNWRWQPTAW showed the highest similarity to residues 352–365 of Fusobacterium periodonticum outer membrane protein A (FomA), which has an additional N-terminal Cys residue. This sequence was identical between F. nucleatum and F. periodonticum except for the sixth residue. The peptide was synthesized using F-moc chemistry and was conjugated to keyhole limpet hemocyanin using maleimidobenzoic acid N-hydroxy succinimide ester, and the antibody was prepared as described previously [32]. To quantify FomA, immunoblotting was performed as described earlier. Ten micrograms of biofilm organized in the lower well, which was monocultured and cocultured with P. gingivalis, was separated using SDS-PAGE and transferred onto a PVDF membrane. FomA was stained with anti-FomA antibody (1:1,000 dilution) and goat anti-rabbit IgG (1:3,000 dilution). The membrane was scanned, and the staining intensity of the developed 39-kDa bands was evaluated using Image-Quant TL software v.8.1 (GE Healthcare, Little Chalfont, UK).
Structural characterization of monoclonal antibodies targeting C-terminal Ser404 region of phosphorylated tau protein
Published in mAbs, 2019
Jessica E. Chukwu, Erin E. Congdon, Einar M. Sigurdsson, Xiang-Peng Kong
MAbs 8B2, 6B2, and 4E6 were generated by GenScript as previously described.11,12 Briefly, wild type BALB/c mice were immunized with a peptide encompassing the pSer396/pSer404 region conjugated to keyhole limpet hemocyanin via a cysteine residue (cTDHGAEIVYK(pS)PVVSGDT(pS)PRHL). Hybridoma fusions were screened by ELISA and 8B2, 6B2, and 4E6 clones were selected based on binding to the phospho-tau immunogen. Although mAbs 4E6 and 6B2 were functionally characterized previously,11,12 mAb 8B2 has never been published before. Peptides used in ELISA and crystallization were synthesized by W.M. Keck Biotechnology Resource Center or GenScript (Table 1). The lyophilized peptides were solubilized in water to a stock concentration of 10 mg/mL before mixing with the Fabs for crystallization.
Prospects of targeted and immune therapies in SCLC
Published in Expert Review of Anticancer Therapy, 2019
Lizza E. L. Hendriks, Jessica Menis, Martin Reck
In a phase I trial (N = 16), vaccination of SCLC patients (LD and ED-SCLC) with a major response to their initial therapy with synthetic fucosyl GM-1 conjugated to keyhole limpet hemocyanin at different dose levels was safe and induced an IgM-antibody response. However, no radiological responses were observed [119]. Another phase I trial vaccinated SCLC patients (N = 18, LD and ED-SCLC) responding to initial therapy, with a polySA vaccine. PolySA is a polymer side chain bound to the neural cell adhesion molecule that is extensively expressed on the surface of SCLC cells. The vaccine was safe and antibody responses were documented but radiological responses were not provided. Median OS was 22.9 months rather encouraging in this patient population [120].