4-Hydroxy-3-nitrophenylacetyl hapten is conjugated to BSA (Bovine Serum Albumin) protein through lysine by amide bonds.
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Background
T-cell-dependent immunization against antigens induces somatic hypermutation in the V region of the antibody, resulting in increased binding affinity, which is known as affinity maturation. Affinity maturation of antibodies is a form of protein evolution in nature. Mutations are introduced either in the direct antigen-contact region or in the indirect region, including the framework region, resulting in changes in antibody structure and structural dynamics. The combination of crystal structure, binding kinetics and biophysical methods such as thermodynamics and folding thermodynamics can provide information on the relationship between protein structure and activity. Mature antibodies return to a stable level similar to that of germline antibodies upon binding to antigen.
Previous studies have shown that immunization with (4-hydroxy-3-nitrophenyl)acetyl (NP) results in the secretion of at least two types of antibodies: one with Tyr at position 95 of the heavy chain and the other with Gly (known as Tyr95 and Gly95 types). The former occurs at an early stage, whereas the latter occurs at a late stage, after secondary immunization, although the Fv domains of these antibodies are encoded by the same genes with variable heavy and light chains. Primary antibodies produced early in immunization have a lower affinity and those produced later have a higher affinity. Single-chain Fv (scFv) is a suitable model for the study of antibody structure and function, and its thermodynamic and kinetic parameters for interaction with various antigens are consistent with those of intact antibodies.
Figure 1. Close-up view of crystal structure of C6 scFv in complex with NP (Source: Oda M. 2022)
Elucidating the structural and physical properties of antibodies that emerge during affinity maturation can also help us to understand the molecular recognition mechanisms of proteins and provide a basis for rational protein design, such as in vitro methods to improve the affinity and specificity of antibodies. For this purpose, researchers have used various hapten systems for their studies, such as NP, choline phosphate and 2-phenyl oxazolone. These studies suggest that dynamic changes in B cell clones are responsible for antibody production during immunization. The structural properties of anti-NP antibodies during affinity maturation have been well analyzed. Germline-type antibodies have antigen-binding affinities as low as 105-106 M-1, whereas affinity-matured antibodies sometimes have antigen-binding affinities as high as 109 M-1. One research group reported that mutation of Trp at position 33 of the heavy chain to Leu resulted in an approximately 10-fold increase in affinity.
Alternative Names
(4-hydroxy-3-nitrophenyl)acetyl [BSA], Ratio 1-9
References
1. Sato Y, et al. Pronounced effect of hapten binding on thermal stability of an anti-(4-hydroxy-3-nitrophenyl)acetyl antibody possessing a glycine residue at position 95 of the heavy chain. Mol Immunol. 2017 May;85:130-136.
2. Oda M. Structural, functional, and physiological properties of anti-(4-hydroxy-3-nitrophenyl)acetyl antibodies during the course of affinity maturation. Biophys Rev. 2022 Oct 20;14(6):1521-1526.
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References
Model robust profile monitoring for the generalized linear mixed model for Phase I analysis
APPLIED STOCHASTIC MODELS IN BUSINESS AND INDUSTRY
Authors: Bandara, Keerthi; Abdel-Salam, Abdel-Salam G.; Birch, Jeffrey B.
The generalized linear mixed model (GLMM) becomes very popular in profile monitoring, especially when the production processes follow nonnormal distribution. In most of the real-life applications in industry, medicine, biology horizontal ellipsis and so on researchers assume that the response variable follows a Bernoulli or Binomial distribution. The majority of previous studies in profile monitoring focused on parametric modeling using the logistic regression model, with both fixed or random effects, under the assumption of correct model specification. This research considers those cases where the parametric logistic regression model for the family of profiles is unknown or at least uncertain. Consequently, we propose two mixed model methods to monitor profiles from the exponential family: a nonparametric (NP) regression method based on the penalized spline regression technique and a semiparametric method (model robust profile monitoring for the generalized linear mixed model) which combines the advantages of both the parametric and NP methods. Several Hotelling T2 charts that have been studied for a binary response variable with replicates for Phase I profile monitoring. The performance of the proposed method is evaluated by using mean squares of errors and probability of signals criteria. The results showed satisfactory performance of the proposed control charts.
Dynamic DAG Scheduling on Multiprocessor Systems: Reliability, Energy, and Makespan
IEEE TRANSACTIONS ON COMPUTER-AIDED DESIGN OF INTEGRATED CIRCUITS AND SYSTEMS
Multiprocessor systems are increasingly deployed in real-time applications, where reliability, energy consumption, and makespan are often the main scheduling objectives. In this work, we investigate the dynamic scheduling of tasks modeled by directed acyclic graphs (DAGs), which is an NP-hard problem with all existing methods being heuristics. Our contributions have two steps: 1) assuming that the allocation of DAG nodes to processors is given, we propose optimal energy allocation (OEA) and search-based OEA (SOEA)-the first optimal methods that minimize the energy consumption while satisfying the reliability requirement-for homogeneous and heterogeneous systems, respectively and 2) we present a novel scheduling algorithm outdegree scheduling (ODS) that allocates the DAG nodes according to their out-degrees, and considering energy consumption, reliability, as well as dynamic finish time. ODS dominates the widely applied heterogeneous earliest finish time (HEFT) in makespan. Combining SOEA with ODS makes a complete solution to the problem of dynamic DAG scheduling on multiprocessor systems, and achieves generally better results compared to the existing approaches. Specifically, in most cases, we are better on all the three objectives, i.e., reliability, energy, as well as makespan, and in other cases, we are better on some of the objectives.
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Figure 1. Close-up view of crystal structure of C6 scFv in complex with NP
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