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KOKONO THE BEACH BOYI SERIESAs Brian entered an extended period of seclusion, the band took a more democratic approach in the studio, resulting in a series of irreverent, eclectic records-epitomized by 1971’s self-effacing Surf’s Up-that were proudly out of step with the dominant acid-rock trends of the day, but whose inspired fusion of soul, psychedelia, and orchestral pop would be later reclaimed by future generations of home-recording indie savants. But Brian’s obsessive tendencies dovetailed with his worsening mental health (spurred by the lingering trauma of an abusive upbringing), resulting in the abandonment of Pet Sounds’ would-be grandiose follow-up, Smile (which was strip-mined for 1967’s loopy psych-pop pastiche Smiley Smile). Brian’s auterist vision-and his increasingly poignant songcraft-achieved peak clarity with 1966’s chamber-pop masterpiece Pet Sounds, the album that inspired The Beatles to venture into Pepperland and heralded rock’s elevation into high art. The Fat Boys & The Beach Boys - Wipeout (Promo) 1987 VHS-Rip. suburb of Hawthorne in 1961, brothers Brian, Carl, and Dennis Wilson, their cousin Mike Love, and high-school pal Al Jardine defined the sunny California fantasy forevermore with wave-riding soundtracks like “Surfin’ U.S.A.” As the surf fad dried up, Brian expanded his primary-songwriter role to become the band’s all-knowing creative director and, on mid-’60s delights like “California Girls,” he refashioned The Beach Boys into the male equivalent of the girl groups ensconced within Phil Spector’s Wall of Sound. But over the next half-century, they’d come to symbolize its divided soul, and the psychic tug-of-war between flag-waving optimism and darker truths. In their early-’60s inception, The Beach Boys were nothing less than the sound of America. Men: 10 x weight (kg) + 6.25 x height (cm) – 5 x age (y) + 5.Our BMR calculator uses three different equations to determine your Basal Metabolic Rate (2). 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(b–d) Schematic representation of certain bsAb formats within the three different groups of bsAbs, namely the asymmetric (b), symmetric (c) and fragment-based bsAbs (d). The major affinity ligand-binding sites are also indicated with an arrow at the respective positions on the IgG. The VH and VL domains make up the variable fragment (Fv) domain. The VL, VH, CL and CH1 domains make up the antigen-binding fragment (Fab), whereas the CH2 and CH3 domains constitute the crystallizable fragment (Fc) region. The HC comprises of VH, CH1, hinge, CH2 and CH3 domains, whereas the LC comprises of VL and CL domains. (a) Schematic representation of an immunoglobulin G (IgG) monoclonal antibody (mAb), which consists of two heavy chains (HCs, dark green) and two light chains (LCs, light green). To this end, this review aims to present the key structural properties of bsAbs and their associated byproducts, outlining the current major purification methods of bsAbs and highlighting the corresponding solutions that have been proposed to circumvent the challenges, as well as to offer a perspective towards future process development. 1) as well as the presence of bsAb-related byproducts, an understanding of which will aid in the identification of potential challenges and therefore design of the optimal strategy for their downstream processing. Although the optimized downstream processing protocols of mAbs serve as a good starting point for the purification of bsAbs, further optimization cannot be fully eliminated due to the differences in their intrinsic structural and concomitant physicochemical properties ( Fig. Many of the current downstream processing methods of bsAbs are built upon the established purification methods of monoclonal antibodies (mAbs), as there are undoubtedly several structural similarities between these antibodies, with the former being derived from at least parts of the latter ( Fig. Yet, in comparison with the numerous detailed reviews outlining the various different formats of bsAbs, along with the associated upstream platform technologies to generate them in order to minimize product-related impurities and their corresponding therapeutic applications, the review of downstream purification of this important class of antibodies is comparatively limited, which may at least in part be attributed to the fewer publications that focus on the purification of these antibodies. The enormous therapeutic potential of bsAbs has led to the development of over 50 different formats of recombinant bsAbs reported so far. INTRODUCTIONīispecific antibodies (bsAbs) demonstrate novel functionalities that yield remarkable promise in improving the drug therapeutic efficacy through the recognition and targeting of two different antigens. Statement of Significance: This review aims to present the key structural properties of bsAbs and their associated byproducts, outlining the current major purification methods of bsAbs and highlighting the corresponding solutions that have been proposed to circumvent the challenges, as well as to offer a perspective towards future process development. Finally, a perspective towards future process development is offered. Here, we outline the current major purification methods of bsAbs, highlighting the corresponding solutions that have been proposed to circumvent the unique challenges presented by this class of antibodies, including differential affinity chromatography, sequential affinity chromatography and the use of salt additives and pH gradients or multistep elutions in various modes of purification. Nevertheless, the downstream processing of bsAbs presents a unique set of challenges due to the presence of bsAb-specific byproducts, such as mispaired products, undesired fragments and higher levels of aggregates, that are otherwise absent or present in lower levels in mAb cell culture supernatants, thus often requiring the design of additional purification strategies in order to obtain products of high purity. Due to the various fundamental structural similarities between bsAbs and monoclonal antibodies (mAbs), many of the current bsAb downstream purification methodologies are based on the established purification processes of mAbs, where affinity, charge, size, hydrophobicity and mixed-mode-based purification are frequently employed. The downstream processing of this class of antibodies is therefore of crucial importance in ensuring that these products can be obtained with high purity and yield. Bispecific antibodies (bsAbs) represent a highly promising class of biotherapeutic modality. |
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