Particle size and zeta potential investigation of synthetic peptide-protein conjugates / Sentetik peptid-protein konjugatlarının parçacık boyutu ve zeta potensiyel analizi

Abstract

Objective: Synthetic peptides are not sufficiently large or complex by themselves to induce immune system because of their small size. Synthetic peptides are usually conjugated to different carriers such as proteins and polyelectrolytes to enhance their immunogenic properties and antigen-specific antibody production (Abs) rate. Thus, the aim of this study is synthesis of peptide-protein covalent conjugates, and size and zeta potential analysis of these conjugates.Methods: In this study, synthetic peptide antigen of the 135-161 amino acids sequence of immunogenic VP1 capsid protein of "A" type foot-and-mouth disease virus (FMDV) was covalently conjugated to bovine serum albumin (BSA) with the carbodiimide method by using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) at different molar ratios of peptide (γ=n Peptide /n BSA ). Particle size and zeta potential analysis of peptide-protein bioconjugates have been performed by using dynamic and electrophoretic light-scattering methods.Results: The size and surface charge of bioconjugates are important factors in a synthetic peptide vaccine. Nevertheless, there virtually no research has been conducted on zeta potential and the size of peptide-protein bioconjugates detailed.Conclusion: Dynamic and electrophoretic light scattering analyses clearly demonstrated that zeta potential of the FMDV 135-161 synthetic peptide-BSA conjugates shifts to less negative potentials and particle sizes increase as the amount of peptide increased in conjugates. The data about peptide-carrier protein conjugates obtained by using these methods are very important for developing peptide-based vaccines.Özet: Amaç: Sentetik peptidler, küçük boyutlarından dolayı, immun sistemin uyarılması için gerekli büyüklükte ve kompleks yapıda değildir. Sentetik peptidler, immunojenik özelliklerinin düzenlenmesi ve antijen-spesifik antikor üretimi oranını arttırmak için genellikle protein ve polielektrolitler gibi farklı taşıyıcılar ile konjuge edilmektedirler.Metod: Bu çalışmada, A tipi şap hastalığı virüsünün (FMDV) VP1 yüzey proteininin 135-161 amino asid dizili antijenik peptidi, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) kullanılarak, sığır serum albumin proteinine kovalent olarak bağlanmıştır. Bağlama peptidin farklı mol oranlarında (γ=n Peptide /n BSA ) gerçekleştirilmiştir. Bulgular: Sentetik peptid aşılarında biyokonjugatların boyut ve yüzey yükleri önemli faktörlerdir. Buna karşın peptid-protein biyokonjugatlarının detaylı olarak boyut ve zeta potansiyel analizlerinin yapıldığı neredeyse hiç çalışma bulunmamaktadır.Sonuç: Dinamik ve elektroforetik ışık saçılması sonuçları; FMDV 135-161 sentetik peptid-BSA konjugatlarının zeta potansiyel değerinin mutlak mutlak küçük değerlere kaydığını ve parçacık boyutunun da peptidin artan oranları ile beraber arttığını göstermiştir. Bu yöntemler kullanı-larak peptid-taşıycı protein konjugatları hakkında elde edilen veriler peptid bazlı aşı geliştirme çalışmalarında oldukça önemlidir.

Quantitative and Label-Free Detection of Protein Kinase A Activity Based on Surface-Enhanced Raman Spectroscopy with Gold Nanostars

“…Kem powder with 95% purity [GenScript (Hong Kong) Limited] was conjugated to BSA by carbodiimide cross-linker chemistry. 26 Briefly, BSA was dissolved in phosphate buffer at pH 5 to facilitate efficient cross-linking. 1-Ethyl-3-(3-(dimethylamino)propyl)-carbodiimide hydrochloride (EDC) powder was then added to BSA at twice the molar ratio of BSA and stirred at 4 °C for 5 min to activate the carboxylic acid groups on BSA.…”

“…Although both BSA and kemptide contained carboxylates and amines, random polymerization of polypeptides caused by EDC-mediated cross-linking could be minimized by a Kem-to-BSA molar ratio of less than 3, 26 where multiple Kem could be conjugated on one BSA without polymerization. The molar concentration of kemptide in purified BSA−Kem was estimated to be twice that of BSA since the initial Kem-to-BSA ratio was 2, with the assumption that all Kem were conjugated to BSA.…”

“…Kem powder with 95% purity [GenScript (Hong Kong) Limited] was conjugated to BSA by carbodiimide cross-linker chemistry . Briefly, BSA was dissolved in phosphate buffer at pH 5 to facilitate efficient cross-linking.…”

Virus-like Particles: Fundamentals and Biomedical Applications

“…In terms of the sample analysis, DLS measurement conditions must be defined to avoid variabilities on the resultant ζ-potential; the factors that influence this value are temperature, solvent ratio (e.g., organic solvents), pH, presence or not of surfactants, the existence of ions in a solution, and ionic strength [ 123 , 124 , 125 ]. In addition, the DLS analysis must consider the intrinsic features of the produced NPs to report the acquired ζ-potential; these characteristics include their size, shape, sedimentation, polydispersity, and presence of conjugates [ 126 , 127 ].…”

Comparative evaluation of hesperetin loaded nanoparticles for anticancer activity against C6 glioma cancer cells

“…The encapsulation efficiency and drug loading were determined according to the following equation. Mean particle size (Z-Ave), zeta potential (f), polydispersity index (PDI) and surface morphology Z-Ave and f values of the HspNPs were measured with photon correlation spectroscopy using a Zetasizer Nano ZS (Malvern Instruments, Worcestershire, UK) [25]. For the purpose of visualizing surface morphology and size distrubition of nanoparticles, field emission scanning electron microscopy was used (JSM-7001FA, JEOL, Tokyo, Japan) with an acceleration voltage of 10 kV [26].…”