1. Adsorption Separation of Immunoglobulin G using Modified Dextran (adapted from Belter, Cussler, & Hu, pg 153).
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1 M 47, ochemcal Processes Fall 007 Homewor #8 Fr. 09 Nov., 007. Adsorpton Separaton o Immunoglobuln G usng Moded Dextran (adapted rom elter, ussler, & Hu, pg 5). The equlbrum between mmunoglobuln G and a moded dextran (the adsorbent) can be descrbed by the angmur sotherm. Dextran wll adsorb up to 7.8x -6 es o mmunoglobuln G per o adsorbent and the angmur constant, K, s.9x -5 es/lter. The adsorbent densty s gram o adsorbent per o adsorbent and the vod racton o a column paced wth ths adsorbent s a) What s the total adsorpton capacty o a column paced wth moded dextran the eed concentraton o mmunoglobuln G s x -5 es/lter? (use column data rom b)) b) What s the mean retenton tme or mmunoglobuln G n a column o dameter equal to 5 wth a eed low rate o lter per mnute, eed concentraton o x -5 es/lter, and a length o meter? c) How many columns would have to be employed to recover mmunoglobuln G rom a eed tan o,000 ter volume assumng the same eed concentraton?. Travel Dstance o Solutes A and n a hromatographc olumn. Problem.6 o the text, parts a and b. Assume that A 0. / and 0.05 / n the lqud phase o the column at equlbrum wth the adsorbed solute.. Determnng Tme to Elute a Solutes A and rom a hromatographc olumn. Problem.7 o the text Due Fr. 6 Nov., 007.
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8 M 47, ochemcal Processes Fall 00 Homewor #7 Soluton Fr. 7 Nov., 00. Determnng Tme to Elute a Solutes A and rom a hromatographc olumn. Problem.7 o the text The undamental eqn. For gel chromatography s V e V o K D V For A: V e 0 0.5(0 ) 5 T A 5 /0 /hr 0.5 hr For A: V e 0 0.5(0 ) 4.5 T A 4.5 /0 /hr 0.45 hr
9 M 47, ochemcal Processes Fall 00 Homewor #8 Fr. 4 Nov., 005. Adsorpton Separaton o Immunoglobuln G usng Moded Dextran (adapted rom elter, ussler, & Hu, pg 5). The equlbrum between mmunoglobuln G and a moded dextran (the adsorbent) can be descrbed by the angmur sotherm. Dextran wll adsorb up to 7.8x -6 es o mmunoglobuln G per o adsorbent and the angmur constant, K, s.9x -5 es/lter. The adsorbent densty s gram o adsorbent per o adsorbent and the vod racton o a column paced wth ths adsorbent s a) What s the total adsorpton capacty o a column paced wth moded dextran the eed concentraton o mmunoglobuln G s x -5 es/lter? (use column data rom b)) b) What s the mean retenton tme or mmunoglobuln G n a column o dameter equal to 5 wth a eed low rate o lter per mnute, eed concentraton o x -5 es/lter, and a length o meter? c) How many columns would have to be employed to recover mmunoglobuln G rom a eed tan o,000 ter volume assumng the same eed concentraton?. Travel Dstance o Solutes A and n a hromatographc olumn. Problem.6 o the text, parts a and b. Assume that A 0. / and 0.05 / n the lqud phase o the column at equlbrum wth the adsorbed solute.. Determnng Tme to Elute a Solutes A and rom a hromatographc olumn. Problem.7 o the text Due Fr. Nov., 005.
10 . Adsorpton Separaton o Immunoglobuln G usng Moded Dextran (adapted rom elter, ussler, and Hu, pg 5) Inormaton angmur sotherm K Dextran wll adsorb up to 7.8E - 6 /IG/^ K S SMAX.9 5 / g ρ adsorbent ( vod racton) Soluton a) Adsorpton capacty o the column D 5 m 0 apacty o 5 the column apacty o the column 5 Volume o the column π S apacty o the column : SMAX ( volume o adsorbent)( soluteconcentraton on adsorbent) ( )( volume o the column)( ) ( 0) ( 0.40)( 96.5 ) S
11 b) Mean retenton tme mn mn mn mn 00 mn 00 mn ADSORENT SOUTION ADSORENT SOUTION ADSORENT SOUTION SMAX SMAX adsorbent g t K K K d d A F U g F U t π ρ ρ c) Number o columns to recover product n columns columns o Number 5
12 . Travel dstance o solutes A and n a chromatographc column. Inormaton c m () c c A, A A 0.5 A 6 0. solute/ lqud 0.0 solute/ lqud mass o adsorbent g 000 ed volume 50 V 50 Soluton a) Poston o each band n the column 0. solute A adsorbed/ adsorbent 0.05 solute adsorbed/ adsorbent
13 m X X M c c c dc d M M A V X adsorbent adsorbent A adsorbent adsorbent For 0.5 A 0. For A column (/) adsorbent per unt volume o the amount o s b) M R M R X X A A A
14 . Determnng Tme to Elute Solutes A and rom a hromatographc olumn Inormaton K 0.5 K V V V 0 DA D 0 0 column F 0 h Soluton Estmate ext tme o A and
15 V0 K DV t F 0 t A 0 h 0 t 0 h ( 0.5)( 0 ) 0.5h ( 0.5)( 0 ) 0.45h
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