Preparation of Humanized Ovarian Carcinoma Anti-Idiotypic Minibody

Abstract

Murine anti-idiotypic monoclonal antibody (MAb) 6B11 mimicking the tumor-associated antigen OC166-9 is used as a vaccine for the induction of an anti-tumoral immunity in experiments of in vitro and in vivo animal model with ovarian carcinoma. In this article, we have humanized 6B11 anti-idiotypic minibody using overlap polymerase chain reaction (PCR) and DNA recombinant technique, prokaryotic expression vector was produced by genetic fusion of 6B11V(L)-V(H) to human IgG1 hinge and CH3 region. Transformed E. coli BL21(DE3) were propagated and induced by isopropyl-beta D-thiogalactopyranoside (IPTG). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that a protein band with molecular weight of 50kD appeared as the expected size after transformation. Molecular weight of 100 kDa may be examined by electrophoresis in nondenaturing systems. The fusion protein was analyzed with enzyme-linked immunosorbant assay (ELISA), inhibition ELISA tests and Western blot, respectively. The humanized anti-idiotype minibody showed capacity of bivalent binding to ovarian cancer MAb COC166-9 and goat anti-human immunoglobulin IgG1. It is useful reagents for clinical use.

Preliminary Safety and Potential Effect of 6B11-OCIK Adoptive Cell Therapy Against Platinum-Resistant Recurrent or Refractory Ovarian Cancer

“…The 6B11 monoclonal antibody mimics the ovarian cancer–associated antigen OC166-9 and induces specific humoral and cellular immunity against ovarian cancer. The humanized modified 6B11 minibody (6B11mini) was obtained by combining the single chain of the 6B11 antibody with the human IgG hinge region ( 16 ). Anti-idiotypic 6B11mini–pulsed DCs were shown to induce T cell responses for specific killing against autologous ovarian cancer ( 17 ).…”

Stochastic Gravitational Wave Background from Global Cosmic Strings

“…where a is cosmic scale factor. For n = 3 (matter domination), 4 (radiation domination), and 6 (kination, to be discussed later), we find C eff 1.32, 2.26, 2.70 at large N , respectively [38].…”

“…Later we will also present results for a log uniform distribution up to α ∼ 1 as suggested in [28] based on simulating the first a few e-folds of Hubble expansion after the formation of a global string network. More comprehensive discussion on the sensitivity to loop distribution and the effect of possible deviation from scaling behavior will be presented in [38].…”

The Strategy of Conditionally Replicating Adenovirus-Mediated PreS2 Mini-Antibody Expression Has Dual Effects of Inhibiting HBV Infection and Preventing Hepatocellular Carcinoma

“…11,12 As a result, mPreS2 might present more effective inhibition of HBV infection and antitumor activity. In addition, mPreS2 did not need glycosylation after translation as a result of the deletion of the CH2 region and was suitable for high-volume production in yeast or E. coli systems, 13 which might create the conditions for translational research and clinical application. Second, we utilized the conditionally replicating adenovirus vector with the human survivin promoter to carry and express mPreS2, aiming to enhance its targeting to cancer cells.…”