Antigen preparations in the form of detergent-solubilized cell lysates could, in principle, render membrane proteins (MPs) compatible with in vitro antibody engineering technologies. To this end, detergent-solubilized cell lysates were coupled with the yeast surface display platform to affinity mature an anti-transferrin receptor (TfR) single-chain antibody (scFv). Lysates were generated from TfR-expressing HEK293 cells by solubilization with detergent-containing buffer after undergoing plasma membrane-restricted biotinylation. Lysate-resident TfR was then combined with a mutagenic anti-TfR scFv library in a competitive, dissociation rate screen, and scFvs were identified with up to 4-fold improved dissociation rates on the surface of yeast. Importantly, although the lysates contained a complex mixture of biotinylated proteins, the engineered scFvs retained their TfR binding specificity. When secreted by yeast as soluble proteins, mutant scFvs bound to cell surface TfR with 3-7-fold improvements in equilibrium binding affinity. Although a known MP antigen was targeted for purposes of this study, employing biotin tagging as a means of antigen detection makes the lysate-based approach particularly flexible. We have previously shown that yeast display can be used to identify lead antibodies using cell lysate-resident MP antigens, and combined with this work showing that antibodies can also be quantitatively engineered using cell lysates, these approaches may provide a high-throughput platform for generation and optimization of antibodies against MPs.
Background Dislocation after primary total hip arthroplasty (THA) has an incidence of 2–3%. Approximately 77% of dislocations occur within the first year after surgery. The SuperPATH technique is a minimally invasive approach for THA that preserves soft tissue attachments. The purpose of this study is to describe the dislocation rate at 1 year after SuperPATH primary THA. Methods All elective primary THAs performed by the senior author using the SuperPATH approach. Exclusion criteria were acute femoral neck fracture, revision surgery, or malignancy. There were 214 of 279 eligible patients available for telephone interviews (76.7%). Medical records were reviewed for secondary outcomes including early and late complications, cup positioning, distance ambulated on postoperative day one, discharge destination, and blood transfusions. Results Mean age at surgery was 64 ± 10.8 years and mean time to telephone follow up was 773 ± 269.7 days. There were 104 female and 110 male patients. There were zero dislocations reported. Blood transfusions were performed in 3.7% of patients, and 75.7% were discharged to home at an average of 2.3 ± 1.0 days. Cup position averaged 43.6 ± 5.2° abduction and 20.9 ± 6.2° anteversion, with an average leg length discrepancy of 3.6 ± 3.32 mm. Complications included three intraoperative calcar fractures, one periprosthetic femur fracture, one early femoral revision, three superficial infections, and one instance of wound necrosis. Conclusion SuperPATH approach is safe for use in primary THA resulting in a low dislocation rate.
Background: Open fractures are debilitating injuries for athletes. No prior studies have investigated open fractures in National Football League (NFL) players. Purpose: To compare outcomes after open fracture in NFL players in terms of (1) time to return to sport (RTS), (2) postinjury career length and games played per season, (3) postinjury performance, and (4) postinjury performance compared with matched controls. Study Design: Retrospective comparative series; Level of evidence, 3. Methods: Publicly available records were used to identify NFL players who had sustained an open fracture between 1970 and 2018. Controls were matched to injured players by age, experience, position, and preinjury performance. RTS was defined as playing in at least 1 NFL game after open fracture. Comparisons between injured and control players were made using the paired-samples Student t test. Results: Injuries in 37 players were analyzed (age, 27.2 ± 3.6 years; experience, 4.4 ± 3.6 seasons). The 3 most common locations for open fracture were the tibia/fibula (n = 16), hand/finger (n = 12), and forearm/wrist (n = 3). A total of 30 (81%) players had a mean time of RTS of 9.3 ± 8.2 months after open fracture; of these players, 4 (13.3%) who sustained hand/finger open fracture did not undergo surgical treatment. There was no difference in postinjury career length or games played per season between control and injured players. Postinjury performance was similar to preinjury performance in injured players, and postinjury performance scores were similar between injured and control players. There were significant differences between players who sustained upper extremity and lower extremity open fractures in RTS time (4.0 ± 4.8 vs 14.6 ± 7.4 months, respectively; P = .00007) and postinjury performance (6.4 ± 4.3 vs 3.3 ± 2.1, respectively; P = .03). Conclusion: RTS after open fracture among NFL players was high. Players who sustained an open fracture had similar games played per season, career length, and performance compared with matched controls. Players who sustained an upper extremity open fracture had a faster RTS time, higher RTS rate, and improved postinjury performance compared with players who sustained a lower extremity open fracture.
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